Stocking density and artificial habitat influence stock structure and yield from intensive nursery systems for mud crabs Scylla serrata (Forsskål 1775)

Intensive nursery systems are designed to culture mud crab postlarvae through a critical phase in preparation for stocking into growout systems. This study investigated the influence of stocking density and provision of artificial habitat on the yield of a cage culture system. For each of three batches of postlarvae, survival, growth and claw loss were assessed after each of three nursery phases ending at crab instars C1/C2, C4/C5 and C7/C8. Survival through the first phase was highly variable among batches with a maximum survival of 80% from megalops to a mean crab instar of 1.5. Stocking density between 625 and 2300 m-2 did not influence survival or growth in this first phase. Stocking densities tested in phases 2 and 3 were 62.5, 125 and 250 m -2. At the end of phases 2 and 3, there were five instar stages present, representing a more than 20-fold size disparity within the populations. Survival became increasingly density-sensitive following the first phase, with higher densities resulting in significantly lower survival (phase 2: 63% vs. 79%; phase 3: 57% vs. 64%). The addition of artificial habitat in the form of pleated netting significantly improved survival at all densities. The mean instar attained by the end of phase 2 was significantly larger at a lower stocking density and without artificial habitat. No significant effect of density or habitat on harvest size was detected in phase 3. The highest incidence of claw loss was 36% but was reduced by lowering stocking densities and addition of habitat. For intensive commercial production, yield can be significantly increased by addition of a simple net structure but rapidly decreases the longer crablets remain in the nursery.

Landscape structure influences tree density patterns in fragmented woodlands in semi-arid eastern Australia.

Landscape and local-scale influences are important drivers of plant community structure. However, their relative contribution and the degree to which they interact remain unclear. We quantified the extent to which landscape structure, within-patch habitat and their confounding effects determine post-clearing tree densities and composition in agricultural landscapes in eastern subtropical Australia. Landscape structure (incorporating habitat fragmentation and loss) and within-patch (site) features were quantified for 60 remnant patches of Eucalyptus populnea (Myrtaceae) woodland. Tree density and species for three ecological maturity classes (regeneration, early maturity, late maturity) and local site features were assessed in one 100 × 10 m plot per patch. All but one landscape characteristic was determined within a 1.3-km radius of plots; Euclidean nearest neighbour distance was measured inside a 5-km radius. Variation in tree density and composition for each maturity class was partitioned into independent landscape, independent site and joint effects of landscape and site features using redundancy analysis. Independent site effects explained more variation in regeneration density and composition than pure landscape effects; significant predictors were the proportion of early and late maturity trees at a site, rainfall and the associated interaction. Conversely, landscape structure explained greater variation in early and late maturity tree density and composition than site predictors. Area of remnant native vegetation within a landscape and patch characteristics (area, shape, edge contrast) were significant predictors of early maturity tree density. However, 31% of the explained variation in early mature tree differences represented confounding influences of landscape and local variables. We suggest that within-patch characteristics are important in influencing semi-arid woodland tree regeneration. However, independent and confounding effects of landscape structure resulting from previous vegetation clearing may have exerted a greater historical influence on older cohorts and should be accounted for when examining woodland dynamics across a broader range of environments.

The stock structure of grey mackerel Scomberomorus semifasciatus in Australia as inferred from its parasite fauna

The scombrid Scomberomorus semifasciatus is an important component of inshore fisheries in tropical Australia. Data on the parasite fauna of 593 fish from areas off northern and eastern Australia were examined for evidence of discrete fish populations. The parasites used were juveniles of Pterobothrium pearsoni, Callitetrarhynchus gracilis, Anisakis simplex (sensu latu) and Terranova sp. Tukey Kramer pairwise comparisons gave significant differences in the abundances of two or more parasites between fish from the east coast, the eastern Gulf of Carpentaria and the remainder of northern Australia. Multivariate analysis gave further evidence of differences and the results suggest that at least 4 populations or stocks of grey mackerel occur along the northern and eastern coastline of Australia.

Measuring and predicting canopy nitrogen nutrition in wheat using a spectral index—The canopy chlorophyll content index (CCCI).

Varying the spatial distribution of applied nitrogen (N) fertilizer to match demand in crops has been shown to increase profits in Australia. Better matching the timing of N inputs to plant requirements has been shown to improve nitrogen use efficiency and crop yields and could reduce nitrous oxide emissions from broad acre grains. Farmers in the wheat production area of south eastern Australia are increasingly splitting N application with the second timing applied at stem elongation (Zadoks 30). Spectral indices have shown the ability to detect crop canopy N status but a robust method using a consistent calibration that functions across seasons has been lacking. One spectral index, the canopy chlorophyll content index (CCCI) designed to detect canopy N using three wavebands along the "red edge" of the spectrum was combined with the canopy nitrogen index (CNI), which was developed to normalize for crop biomass and correct for the N dilution effect of crop canopies. The CCCI-CNI index approach was applied to a 3-year study to develop a single calibration derived from a wheat crop sown in research plots near Horsham, Victoria, Australia. The index was able to predict canopy N (g m-2) from Zadoks 14-37 with an r2 of 0.97 and RMSE of 0.65 g N m-2 when dry weight biomass by area was also considered. We suggest that measures of N estimated from remote methods use N per unit area as the metric and that reference directly to canopy %N is not an appropriate method for estimating plant concentration without first accounting for the N dilution effect. This approach provides a link to crop development rather than creating a purely numerical relationship. The sole biophysical input, biomass, is challenging to quantify robustly via spectral methods. Combining remote sensing with crop modelling could provide a robust method for estimating biomass and therefore a method to estimate canopy N remotely. Future research will explore this and the use of active and passive sensor technologies for use in precision farming for targeted N management.

Understanding invasion history: genetic structure and diversity of two globally invasive plants and implications for their management

Aim: Resolving the origin of invasive plant species is important for understanding the introduction histories of successful invaders and aiding strategies aimed at their management. This study aimed to infer the number and origin(s) of introduction for the globally invasive species, Macfadyena unguis-cati and Jatropha gossypiifolia using molecular data. Location: Native range: Neotropics; Invaded range: North America, Africa, Europe, Asia, Pacific Islands and Australia. Methods: We used chloroplast microsatellites (cpSSRs) to elucidate the origin(s) of introduced populations and calculated the genetic diversity in native and introduced regions. Results: Strong genetic structure was found within the native range of M. unguis-cati, but no genetic structuring was evident in the native range of J. gossypiifolia. Overall, 27 haplotypes were found in the native range of M. unguis-cati. Only four haplotypes were found in the introduced range, with more than 96% of introduced specimens matching a haplotype from Paraguay. In contrast, 15 haplotypes were found in the introduced range of J. gossypiifolia, with all invasive populations, except New Caledonia, comprising multiple haplotypes. Main conclusions: These data show that two invasive plant species from the same native range have had vastly different introduction histories in their non-native ranges. Invasive populations of M. unguis-cati probably came from a single or few independent introductions, whereas most invasive J. gossypiifolia populations arose from multiple introductions or alternatively from a representative sample of genetic diversity from a panmictic native range. As introduced M. unguis-cati populations are dominated by a single haplotype, locally adapted natural enemies should make the best control agents. However, invasive populations of J. gossypiifolia are genetically diverse and the selection of bio-control agents will be considerably more complex.

Population structure of South African and Australian Pyrenophora teres isolates.

There are two recognized forms of the disease net blotch of barley: the net form caused by Pyrenophora teres f. teres (PTT) and the spot form caused by P. teres f. maculata (PTM). In this study, amplified fragment length polymorphism analysis was used to investigate the genetic diversity and population structure of 60 PTT and 64 PTM isolates collected across Australia (66 isolates) and in the south-western Cape of South Africa (58 isolates). For comparison, P. tritici-repentis, Exserohilum rostratum and Bipolaris sorokiniana samples were also included in the analyses. Both distance-and model-based cluster analyses separated the PTT and PTM isolates into two strongly divergent genetic groups. Significant variation was observed both among the South African and Australian populations of PTT and PTM and among sampling locations for the PTT samples. Results suggest that sexual reproduction between the two forms is unlikely and that reproduction within the PTT and PTM groups occurs mainly asexually.

Temporal genetic structure patterns in tropical maize populations under reciprocal recurrent selection.

In order to investigate the effect of long term recurrent selection on the pattern of gene diversity, thirty randomly-selected individuals from the progenitors (p) and four selection cycles (C0, C3, C6 and C11) were sampled for DNA analysis from the tropical maize (Zea mays L.) breeding populations, Atherton 1 (AT1) and Atherton 2 (AT2). Fifteen polymorphic Simple Sequence Repeat markers amplified a total of 284 and 257 alleles in AT1 and AT2 populations, respectively. Reductions in the number of alleles were observed at advanced selection cycles. About 11 and 12% of the alleles in AT1 and AT2 populations respectively, were near to fixation. However, a higher number of alleles (37% in AT1 and 33% in AT2) were close to extinction. Fisher's exact test and analysis of molecular variance (AMOVA) showed significant population differentiations. Gene diversity estimates and AMOVA revealed increased genetic differentiations at the expense of loss of heterozygosity. Population differentiations were mainly due to fixation of complementary alleles at a locus in the two breeding populations. The estimates of effective population at an advanced selection cycle were close to the population size predicted by the breeding method.

Stock structure of Grey Mackerel, Scomberomorus semifasciatus (Pisces: Scombridae) across northern Australia, based on otolith stable isotope chemistry

The stable isotopes of delta O-18 and delta C-13 in sagittal otolith carbonates were used to determine the stock structure of Grey Mackerel, Scomberomorus semifasciatus. Otoliths were collected from Grey Mackerel at ten locations representing much of their distributional and fisheries range across northern Australia from 2005 to 2007. Across this broad range (similar to 6500 km), fish from four broad locations-Western Australia (S1), Northern Territory and Gulf of Carpentaria (S2, S3, S4, S5, S6, S7), Queensland east coast mid and north sites (S8, S9) and Queensland east coast south site (S10)-had stable isotope values that were significantly different indicating stock separation. Otolith stable isotopes differed more between locations than among years within a location, indicating temporal stability across years. The spatial separation of these populations indicates a complex stock structure across northern Australia. Stocks of S. semifasciatus appear to be associated with large coastal embayments. These results indicate that optimal fisheries management may require a review of the current spatial arrangements, particularly in relation to the evidence of shared stocks in the Gulf of Carpentaria. Furthermore, as the population of S. semifasciatus in Western Australia exhibited high spatial separation from those at all the other locations examined, further research activities should focus on investigating additional locations within Western Australia for an enhanced determination of stock delineation. From the issue entitled "Proceedings of the 4th International Otolith Symposium, 24-28 August 2009, Monterey, California"

Population structure, effective population size and adverse effects of stocking in the endangered Australian eastern freshwater cod Maccullochella ikei

Microsatellite markers were used to examine spatio-temporal genetic variation in the endangered eastern freshwater cod Maccullochella ikei in the Clarence River system, eastern Australia. High levels of population structure were detected. A model-based clustering analysis of multilocus genotypes identified four populations that were highly differentiated by F-statistics (FST = 0· 09 − 0· 49; P < 0· 05), suggesting fragmentation and restricted dispersal particularly among upstream sites. Hatchery breeding programmes were used to re-establish locally extirpated populations and to supplement remnant populations. Bayesian and frequency-based analyses of hatchery fingerling samples provided evidence for population admixture in the hatchery, with the majority of parental stock sourced from distinct upstream sites. Comparison between historical and contemporary wild-caught samples showed a significant loss of heterozygosity (21%) and allelic richness (24%) in the Mann and Nymboida Rivers since the commencement of stocking. Fragmentation may have been a causative factor; however, temporal shifts in allele frequencies suggest swamping with hatchery-produced M. ikei has contributed to the genetic decline in the largest wild population. This study demonstrates the importance of using information on genetic variation and population structure in the management of breeding and stocking programmes, particularly for threatened species.

Negligible evidence for regional genetic population structure for two shark species Rhizoprionodon acutus (Rüppell, 1837) and Sphyrna lewini (Griffith & Smith, 1834) with contrasting biology

Biodiversity of sharks in the tropical Indo-Pacific is high, but species-specific information to assist sustainable resource exploitation is scarce. The null hypothesis of population genetic homogeneity was tested for scalloped hammerhead shark (Sphyrna lewini, n = 237) and the milk shark (Rhizoprionodon acutus, n = 207) from northern and eastern Australia, using nuclear (S. lewini, eight microsatellite loci; R. acutus, six loci) and mitochondrial gene markers (873 base pairs of NADH dehydrogenase subunit 4). We were unable to reject genetic homogeneity for S. lewini, which was as expected based on previous studies of this species. Less expected were similar results for R. acutus, which is more benthic and less vagile than S. lewini. These features are probably driving the genetic break found between Australian and central Indonesian R. acutus (F-statistics; mtDNA, 0.751–0.903, respectively; microsatellite loci, 0.038–0.047 respectively). Our results support the spatially homogeneous monitoring and management plan for shark species in Queensland, Australia.

Evidence for extensive population structure in the white-spotted eagle ray within the Indo-Pacific inferred from mitochondrial gene sequences

The white-spotted eagle ray Aetobatus narinari is a species complex that occurs circumglobally throughout warm-temperate waters. Aetobatus narinari is semi-pelagic and large (up to 300 cm disc width), suggesting high dispersal capabilities and gene flow on a wide spatial scale. Sequence data from two mitochondrial genes, cytochrome b (cytb) and NADH dehydrogenase subunit 4 (ND4), were used to determine the genetic variability within and among 18 sampling locations in the central Indo-Pacific biogeographical region. Populations in the Indo-Pacific were highly genetically structured with c. 70% of the total genetic variation found among three geographical regions (East China Sea, Southeast Asia and Australia). FST was 0.64 for cytb and 0.53 for ND4, with φST values being even larger, that is, 0.78 for cytb and 0.65 for ND4. This high-level genetic partitioning provides strong evidence against extensive gene flow in A. narinari. The degree of genetic population structuring in the Indo-Pacific was similar to that found on a global scale. Global FST was 0.63 for cytb and 0.57 for ND4, and global φST values were 0.94 for cytb and 0.82 for ND4. This suggests that the A. narinari complex may be more speciose than the two or three species proposed to date. Further sampling and genetic analyses are likely to uncover the ‘evolutionarily significant’ and ‘management’ units that are critical to determine the susceptibilities of individual populations to regional fishing pressures and to provide advice on management options. Network analyses showed a close genetic relationship between haplotypes from the central Indo-Pacific and South Africa, providing support for a proposed dispersal pathway from the possible centre of origin of the A. narinari species complex in the Indo-Pacific into the Atlantic Ocean.

IUCN classification zones concord with, but underestimate, the population genetic structure of the zebra shark Stegostoma fasciatum in the Indo-West Pacific

The Indo-West Pacific (IWP), from South Africa in the western Indian Ocean to the western Pacific Ocean, contains some of the most biologically diverse marine habitats on earth, including the greatest biodiversity of chondrichthyan fishes. The region encompasses various densities of human habitation leading to contrasts in the levels of exploitation experienced by chondrichthyans, which are targeted for local consumption and export. The demersal chondrichthyan, the zebra shark, Stegostoma fasciatum, is endemic to the IWP and has two current regional International Union for the Conservation of Nature (IUCN) Red List classifications that reflect differing levels of exploitation: ‘Least Concern’ and ‘Vulnerable’. In this study, we employed mitochondrial ND4 sequence data and 13 microsatellite loci to investigate the population genetic structure of 180 zebra sharks from 13 locations throughout the IWP to test the concordance of IUCN zones with demographic units that have conservation value. Mitochondrial and microsatellite data sets from samples collected throughout northern Australia and Southeast Asia concord with the regional IUCN classifications. However, we found evidence of genetic subdivision within these regions, including subdivision between locations connected by habitat suitable for migration. Furthermore, parametric FST analyses and Bayesian clustering analyses indicated that the primary genetic break within the IWP is not represented by the IUCN classifications but rather is congruent with the Indonesian throughflow current. Our findings indicate that recruitment to areas of high exploitation from nearby healthy populations in zebra sharks is likely to be minimal, and that severe localized depletions are predicted to occur in zebra shark populations throughout the IWP region.

Population genetic structure and genetic diversity of three critically endangered Pristis sawfishes in Australian waters.

Northern Australia is considered to be one of the last strongholds for three critically endangered sawfishes, Pristis zijsron, Pristis clavata, and Pristis microdon, making these populations of global significance. Population structure and levels of genetic diversity were assessed for each species across northern Australia using a portion of the mitochondrial control region. Statistically significant genetic structure was detected in all three species, although it was higher in P. microdon (F-ST = 0.811; N = 149) than in either P. clavata (F-ST = 0.419; N = 73) or P. zijsron (F-ST = 0.202; N = 49), possibly due to a much higher and/or localized level of female philopatry in P. microdon. The overall levels of haplotype diversity in P. zijsron (h = 0.555), P. clavata (h = 0.489), and P. microdon (h = 0.650) were moderate, although it appears to be reduced in the assemblages of P. zijsron and P. clavata in the Gulf of Carpentaria (h = 0.342 and h = 0.083, respectively). Since female migration (replenishment) between regions is unlikely, conservation plans should strive to maintain current levels of diversity and abundances in the regional assemblages of each species.

Stock structure of exploited shark species in north-eastern Australia

The project has provided management and other stakeholders with information necessary to make informed decisions about the management of four of the key exploited shark species caught in the Queensland inshore net fishery and northern New South Wales line fishery. The project has determined that spatial management of milk sharks within Queensland, and scalloped hammerhead, common black tip and Australian black tip sharks within Queensland and New South Wales is appropriate. The project has determined that both black tip shark species are likely to require co-operative management arrangements between Queensland and New South Wales. For scalloped hammerheads separate stocks between the two jurisdictions were identified from the fisheriesdependent samples, however genetic exchange across borders is likely to be facilitated by movement of adult females and perhaps larger males to a lesser extent. This information will greatly assist compliance with the Commonwealth Environment Protection and Biodiversity Conservation Act (1999) for shark fisheries in north-eastern Australia by providing the necessary basis for robust assessment of the status of stocks of the study species, thereby helping to deliver their sustainable harvest. It also helps to achieve objectives of the Australian National Shark Plan. The project provides the appropriate spatial framework for future monitoring and assessment of the study species. This is at a time when shark fisheries are receiving close attention from all sectors and when monitoring programs are being implemented, aimed at better assessment of stock status. This project has provided the crucial information for developing an appropriate monitoring design as well as the necessary basis for making statements about stock status. The project has addressed research priorities identified by the Queensland Fisheries Research Advisory Board, Great Barrier Reef Marine Park Authority and Queensland Fisheries. Previously management has assumed a single stock for each species on the east coast of Queensland, and management of shark fisheries in New South Wales (NSW) and Queensland has been independent of one another. The project has been able to enhance and develop links between research, management and industry. Strong positive relationships with commercial fishers were crucial in the collection of samples throughout the study area and fisheries managers were part of the project team throughout the study period. During the project the study area was extended to include both Queensland and NSW waters, creating mutualistic and positive links between the States’ research and management agencies. Extension of project results included management representatives from NSW and Queensland, as well as the Northern Territory where similar shark fisheries operate and similar species are targeted. The project was able to provide significant human capital development opportunities providing considerable value to the project outcomes. Use of vertebral microchemistry and life history characteristics as stock determination methods provided material for two PhD students based at James Cook University: Ron Schroeder, vertebral chemistry; and Alastair Harry, life history characteristic. The project has developed novel research methods that have great capacity for future application, including: • Development of a simple and rapid genetic diagnostic tool (RT-HRM-PCR assay) for differentiating among the black tip shark species, for which no simple morphological identifier exists; and • Development of laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS) methods for analysing and interpreting microchemical composition of shark vertebrae. The study has provided further confirmation of the effectiveness of using a holistic approach in stock structure studies and justifies investment into such studies.

Characterising population structure and diversity of Australian Eimeria

Develop microsatellite markers to distinguish strains of Eimeria acervulina, E. brunetti and E. maxima. Conduct nationwide sampling of chicken faeces to build baseline of Eimeria population genetic diversity for 5 economically important speces (3 species above plus E. tenella and E. necatrix). Conduct focused local screening to assess temporal changes in populations historically sampled.