Rapid microsatellite marker development for African mahogany ( Khaya senegalensis, Meliaceae) using next-generation sequencing and assessment of its intra-specific genetic diversity.

Khaya senegalensis (African mahogany or dry-zone mahogany) is a high-value hardwood timber species with great potential for forest plantations in northern Australia. The species is distributed across the sub-Saharan belt from Senegal to Sudan and Uganda. Because of heavy exploitation and constraints on natural regeneration and sustainable planting, it is now classified as a vulnerable species. Here, we describe the development of microsatellite markers for K. senegalensis using next-generation sequencing to assess its intra-specific diversity across its natural range, which is a key for successful breeding programs and effective conservation management of the species. Next-generation sequencing yielded 93943 sequences with an average read length of 234bp. The assembled sequences contained 1030 simple sequence repeats, with primers designed for 522 microsatellite loci. Twenty-one microsatellite loci were tested with 11 showing reliable amplification and polymorphism in K. senegalensis. The 11 novel microsatellites, together with one previously published, were used to assess 73 accessions belonging to the Australian K. senegalensis domestication program, sampled from across the natural range of the species. STRUCTURE analysis shows two major clusters, one comprising mainly accessions from west Africa (Senegal to Benin) and the second based in the far eastern limits of the range in Sudan and Uganda. Higher levels of genetic diversity were found in material from western Africa. This suggests that new seed collections from this region may yield more diverse genotypes than those originating from Sudan and Uganda in eastern Africa.

Isolation and characterization of 21 polymorphic microsatellite loci in the iconic Australian lungfish, Neoceratodus forsteri, using the Ion Torrent next-generation sequencing platform

We isolated and characterized 21 microsatellite loci in the vulnerable and iconic Australian lungfish, Neoceratodus forsteri. Loci were screened across eight individuals from the Burnett River and 40 individuals from the Pine River. Genetic diversity was low with between one and six alleles per locus within populations and a maximum expected heterozygosity of 0.774. These loci will now be available to assess effective population sizes and genetic structure in N. forsteri across its natural range in South East Queensland, Australia.

Pleopod Cuticular Morphology as an Index of Moult Stage in the Ornate Rock Lobster, Panulirus ornatus (Fabricius 1789)

Seven discrete stages and substages of moulting in the ornate rock lobster, Panulirus ornatus, have been distinguished by microscopic examination of the cuticle and setae of the pleopods . The diagnostic features and the duration of each of the stages are described. Freezing did not visually alter the tissue features used to identify each moult stage. Pleopod morphology can reliably indicate whether a lobster has moulted within the previous 24 h or is within 72 h of the next ecdysis.

Sustaining productivity of a Vertosol at Warra, Queensland, with fertilisers, no-tillage or legumes. 8. Effect of duration of lucerne ley on soil nitrogen and water, wheat yield and protein.

Soil nitrogen (N) supply in the Vertosols of southern Queensland, Australia has steadily declined as a result of long-term cereal cropping without N fertiliser application or rotations with legumes. Nitrogen-fixing legumes such as lucerne may enhance soil N supply and therefore could be used in lucerne-wheat rotations. However, lucerne leys in this subtropical environment can create a soil moisture deficit, which may persist for a number of seasons. Therefore, we evaluated the effect of varying the duration of a lucerne ley (for up to 4 years) on soil N increase, N supply to wheat, soil water changes, wheat yields and wheat protein on a fertility-depleted Vertosol in a field experiment between 1989 and 1996 at Warra (26degrees 47'S, 150degrees53'E), southern Queensland. The experiment consisted of a wheat-wheat rotation, and 8 treatments of lucerne leys starting in 1989 (phase 1) or 1990 (phase 2) for 1,2,3 or 4 years duration, followed by wheat cropping. Lucerne DM yield and N yield increased with increasing duration of lucerne leys. Soil N increased over time following 2 years of lucerne but there was no further significant increase after 3 or 4 years of lucerne ley. Soil nitrate concentrations increased significantly with all lucerne leys and moved progressively downward in the soil profile from 1992 to 1995. Soil water, especially at 0.9-1.2 m depth, remained significantly lower for the next 3 years after the termination of the 4 year lucerne ley than under continuous wheat. No significant increase in wheat yields was observed from 1992 to 1995, irrespective of the lucerne ley. However, wheat grain protein concentrations were significantly higher under lucerne-wheat than under wheat wheat rotations for 3-5 years. The lucerne yield and soil water and nitrate-N concentrations were satisfactorily simulated with the APSIM model. Although significant N accretion occurred in the soil following lucerne leys, in drier seasons, recharge of the drier soil profile following long duration lucerne occurred after 3 years. Consequently, 3- and 4-year lucerne-wheat rotations resulted in more variable wheat yields than wheat-wheat rotations in this region. The remaining challenge in using lucerne-wheat rotations is balancing the N accretion benefits with plant-available water deficits, which are most likely to occur in the highly variable rainfall conditions of this region.

Fusarium wilt of cotton: Population diversity and implications for management

Fusarium wilt of cotton, caused by the fungus Fusarium oxysporum Schlechtend. f. sp. vasinfectum (Atk.) Snyd. & Hans, was first identified in 1892 in cotton growing in sandy acid soils in Alabama (8). Although the disease was soon discovered in other major cotton-producing areas, it did not become global until the end of the next century. After its original discovery, Fusarium wilt of cotton was reported in Egypt (1902) (30), India (1908) (60), Tanzania (1954) (110), California (1959) (33), Sudan (1960) (44), Israel (1970) (27), Brazil (1978) (5), China (1981) (17), and Australia (1993) (56). In addition to a worldwide distribution, Fusarium wilt occurs in all four of the domesticated cottons, Gossypium arboretum L., G. barbadense L., G. herbaceum L., and G. hirsutum L. (4,30). Disease losses in cotton are highly variable within a country or region. In severely infested fields planted with susceptible cultivars, yield losses can be high. In California, complete crop losses in individual fields have been observed (R. M. Davis, unpublished). Disease loss estimates prepared by the National Cotton Disease Council indicate losses of over 109,000 bales (227 kg or 500 lb) in the United States in 2004 (12).

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.

Optimal management of agricultural systems

To remain competitive, many agricultural systems are now being run along business lines. Systems methodologies are being incorporated, and here evolutionary computation is a valuable tool for identifying more profitable or sustainable solutions. However, agricultural models typically pose some of the more challenging problems for optimisation. This chapter outlines these problems, and then presents a series of three case studies demonstrating how they can be overcome in practice. Firstly, increasingly complex models of Australian livestock enterprises show that evolutionary computation is the only viable optimisation method for these large and difficult problems. On-going research is taking a notably efficient and robust variant, differential evolution, out into real-world systems. Next, models of cropping systems in Australia demonstrate the challenge of dealing with competing objectives, namely maximising farm profit whilst minimising resource degradation. Pareto methods are used to illustrate this trade-off, and these results have proved to be most useful for farm managers in this industry. Finally, land-use planning in the Netherlands demonstrates the size and spatial complexity of real-world problems. Here, GIS-based optimisation techniques are integrated with Pareto methods, producing better solutions which were acceptable to the competing organizations. These three studies all show that evolutionary computation remains the only feasible method for the optimisation of large, complex agricultural problems. An extra benefit is that the resultant population of candidate solutions illustrates trade-offs, and this leads to more informed discussions and better education of the industry decision-makers.

Combined treatments of spinosad and chlorpyrifos-methyl for management of resistant psocid pests (Psocoptera: Liposcelididae) of stored grain

The combined efficacy of spinosad and chlorpyrifos-methyl was determined against four storage psocid pests belonging to genus Liposcelis. This research was undertaken because of the increasing importance of these psocids in stored grain and the problem of finding grain protectants to control resistant strains. Firstly, mortality and reproduction were determined for adults exposed to wheat freshly treated with either spinosad (0.5 and 1 mg kg-1) or chlorpyrifos-methyl (2.5, 5 and 10 mg kg-1) or combinations of spinosad and chlorpyrifos-methyl at 30°C and 70% RH. There were significant effects of application rate of spinosad and chlorpyrifos-methyl, both individually and in combination, on adult mortality and progeny reduction of all four psocids. Liposcelis bostrychophila Badonnel and L. decolor (Pearman) responded similarly, with incomplete control of adults and progeny at both doses of spinosad but complete control in all chlorpyrifos-methyl and combined treatments. In L. entomophila (Enderlein) and L. paeta Pearman, however, complete control of adults and progeny was only achieved in the combined treatments, with the exception of spinosad 0.5 mg kg-1 plus chlorpyrifos-methyl 2.5 mg kg-1 against L. entomophila. Next, combinations of spinosad (0.5 and 1 mg kg-1) and chlorpyrifos-methyl (2.5, 5 and 10 mg kg-1) in bioassays after 0, 1.5 and 3 months storage of treated wheat were evaluated. The best treatment was 1 mg kg -1 of spinosad plus 10 mg kg-1 of chlorpyrifos-methyl, providing up to 3 months of protection against infestations of all four Liposcelis spp. on wheat.

Reducing the short-term mortality of juvenile school prawns (Metapenaeus macleayi) discarded during trawling

A field experiment was carried out in southeastern Australia to assess the short-term mortality and stress incurred by juvenile school prawns (Metapenaeus macleayi) discarded from an estuarine trawler. Some 35% of the prawns died up to 72 h after being caught in a trawl, exposed to air during sorting and separation from the retained catch (as per normal commercial procedures), then discarded into replicate cages. Total mortality was partitioned into that caused by trawling (about 16% of mortalities), and by subsequent sorting and grading (about 19%). Assuming that the majority of the non-penaeid bycatch is excluded from trawls (by the use of bycatch reduction devices), the latter mortalities could be almost eliminated by sorting and separating unwanted school prawns in water-filled compartments. Emersion stress was measured as concentrations of l-lactate in the haemolymph, which were elevated for at least 40 min following capture, but similar among all trawled treatments. l-lactate levels decreased within the first 24 h post-capture, then remained constant over at least the next 48 h, and were greater than baseline levels. The potential benefits associated with subtle changes to handling practices onboard estuarine trawlers are discussed.

Inter-seasonal population dynamics and pest status of Bemisia tabaci (Gennadius) biotype B in an Australian cropping system

Bemisia tabaci, biotype B, commonly known as the silverleaf whitefly (SLW) is an alien species that invaded Australia in the mid-90s. This paper reports on the invasion ecology of SLW and the factors that are likely to have contributed to the first outbreak of this major pest in an Australian cotton cropping system, population dynamics of SLW within whitefly-susceptible crop (cotton and cucurbit) and non-crop vegetation (sowthistle, Sonchus spp.) components of the cropping system were investigated over four consecutive growing seasons (September-June) 2001/02-2004/05 in the Emerald Irrigation Area (EIA) of Queensland, Australia. Based on fixed geo-referenced sampling sites, variation in spatial and temporal abundance of SLW within each system component was quantified to provide baseline data for the development of ecologically sustainable pest management strategies. Parasitism of large (3rd and 4th instars) SLW nymphs by native aphelinid wasps was quantified to determine the potential for natural control of SLW populations. Following the initial outbreak in 2001/02, SLW abundance declined and stabilised over the next three seasons. The population dynamics of SLW is characterised by inter-seasonal population cycling between the non-crop (weed) and cotton components of the EIA cropping system. Cotton was the largest sink for and source of SLW during the study period. Over-wintering populations dispersed from weed host plant sources to cotton in spring followed by a reverse dispersal in late summer and autumn to broad-leaved crops and weeds. A basic spatial source-sink analysis showed that SLW adult and nymph densities were higher in cotton fields that were closer to over-wintering weed sources throughout spring than in fields that were further away. Cucurbit fields were not significant sources of SLW and did not appear to contribute significantly to the regional population dynamics of the pest. Substantial parasitism of nymphal stages throughout the study period indicates that native parasitoid species and other natural enemies are important sources of SLW mortality in Australian cotton production systems. Weather conditions and use of broad-spectrum insecticides for pest control are implicated in the initial outbreak and on-going pest status of SLW in the region.

Potential for using through-boring to improve groundline treatment of Australian wood species: A preliminary study.

An aging electricity distribution system and reduced availability of naturally durable tropical hardwoods in Australia will combine in the next decade to produce a major shortage of poles. One approach to mitigating this shortage is to utilize lower durability species and improve the penetration of preservatives into the refractory heartwood by introducing additional pretreatment processes. A potential method for improving preservative penetration in the critical ground-line zone is through-boring. This process, in which holes are drilled through the pole perpendicular to the grain in the ground-line zone, is widely used in the western United States for treatment of Douglas-fir and may be Suitable for many Australian wood species. The potential for improving heartwood penetration in eucalypts with alkaline-copper-quaternary (ACQ) compound was assessed on heartwood specimens from four species (Eucalyptus cloeziana F.Muell., E. grandis W.Hill ex Maiden, E. obliqua L'Her. and E. pellita F.Muell.) and Lophostemon confertus (R.Br.) Peter G.Wilson & J.T.Wateril). Longitudinal ACQ penetration was extremely shallow in L. confertus and only slightly better in E. cloeziana. Longitudinal penetration was good in both E. obliqua and E. pellita, although there was some variation in treatment results with length of pressure period. The results suggest that through-boring might be a reasonable approach for achieving heartwood penetration in some Eucalyptus species, although further studies are required to assess additional treatment schedules and to determine the effects of the process oil flexural properties of the poles.

Potential of piperonyl butoxide-synergised pyrethrins against psocids (Psocoptera: Liposcelididae) for stored-grain protection.

BACKGROUND: Piperonyl butoxide (PB)-synergised natural pyrethrins (pyrethrin:PB ratio 1:4) were evaluated both as a grain protectant and a disinfestant against four Liposcelidid psocids: Liposcelis bostrychophila Badonnel, L. entomophila (Enderlein), L. decolor (Pearman) and L. paeta Pearman. These are key storage pests in Australia that are difficult to control with the registered grain protectants and are increasingly being reported as pests of stored products in other countries. Firstly, mortality and reproduction of adults were determined in wheat freshly treated at 0.0, 0.75, 1.5, 3 and 6 mg kg-1 of pyrethrins + PB (1:4) at 301C and 702% RH. Next, wheat treated at 0.0, 1.5, 3 and 6 mg kg-1 of pyrethrins + PB (1:4) was stored at 301C and 702% RH and mortality and reproduction of psocids were assessed after 0, 1.5, 3 and 4.5 months of storage. Finally, the potential of synergised pyrethrins as a disinfestant was assessed by establishing time to endpoint mortality for adult psocids exposed to wheat treated at 3 and 6 mg kg-1 of synergised pyrethrins after 0, 3, 6, 9 and 12 h of exposure. RESULTS: Synergised pyrethrins at 6 mg kg-1 provided 3 months of protection against all four Liposcelis spp., and at this rate complete adult mortality of these psocids can be achieved within 6 h of exposure. CONCLUSION: Piperonyl butoxide-synergised pyrethrins have excellent potential both as a grain protectant and as a disinfestant against Liposcelidid.

MOE and MOR assessment technologies for improving graded recovery of exotic pines in Australia.

This project was designed to provide the structural softwood processing industry with the basis for improved green and dry grading to allow maximise MGP grade yields, consistent product performance and reduced processing costs. To achieve this, advanced statistical techniques were used in conjunction with state-of-the-art property measurement systems. Specifically, the project aimed to make two significant steps forward for the Australian structural softwood industry: • assessment of technologies, both existing and novel, that may lead to selection of a consistent, reliable and accurate device for the log yard and green mill. The purpose is to more accurately identify and reject material that will not make a minimum grade of MGP10 downstream; • improved correlation of grading MOE and MOR parameters in the dry mill using new analytical methods and a combination of devices. The three populations tested were stiffness-limited radiata pine, strength-limited radiata pine and Caribbean pine. Resonance tests were conducted on logs prior to sawmilling, and on boards. Raw data from existing in-line systems were captured for the green and dry boards. The dataset was analysed using classical and advanced statistical tools to provide correlations between data sets and to develop efficient strength and stiffness prediction equations. Stiffness and strength prediction algorithms were developed from raw and combined parameters. Parameters were analysed for comparison of prediction capabilities using in-line parameters, off-line parameters and a combination of in-line and off-line parameters. The results show that acoustic resonance techniques have potential for log assessment, to sort for low stiffness and/or low strength, depending on the resource. From the log measurements, a strong correlation was found between the average static MOE of the dried boards within a log and the predicted value. These results have application in segregating logs into structural and non-structural uses. Some commercial technologies are already available for this application such as Hitman LG640. For green boards it was found that in-line and laboratory acoustic devices can provide a good prediction of dry static MOE and moderate prediction for MOR.There is high potential for segregating boards at this stage of processing. Grading after the log breakdown can improve significantly the effectiveness of the mill. Subsequently, reductions in non-structural volumes can be achieved. Depending on the resource it can be expected that a 5 to 8 % reduction in non structural boards won’t be dried with an associated saving of $70 to 85/m3. For dry boards, vibration and a standard Metriguard CLT/HCLT provided a similar level of prediction on stiffness limited resource. However, Metriguard provides a better strength prediction in strength limited resources (due to this equipment’s ability to measure local characteristics). The combination of grading equipment specifically for stiffness related predictors (Metriguard or vibration) with defect detection systems (optical or X-ray scanner) provides a higher level of prediction, especially for MOR. Several commercial technologies are already available for acoustic grading on board such those from Microtec, Luxscan, Falcon engineering or Dynalyse AB for example. Differing combinations of equipment, and their strategic location within the processing chain, can dramatically improve the efficiency of the mill, the level of which will vary depending of the resource. For example, an initial acoustic sorting on green boards combined with an optical scanner associated with an acoustic system for grading dry board can result in a large reduction of the proportion of low value low non-structural produced. The application of classical MLR on several predictors proved to be effective, in particular for MOR predictions. However, the usage of a modern statistics approach(chemometrics tools) such as PLS proved to be more efficient for improving the level of prediction. Compared to existing technologies, the results of the project indicate a good improvement potential for grading in the green mill, ahead of kiln drying and subsequent cost-adding processes. The next stage is the development and refinement of systems for this purpose.

Testing the effectiveness of the SKIM foam fractionator to reduce nutrient levels in prawn farm effluent

To experimentally investigate the effect of the “SKIM” mechanical foam fractionator on suspended material and the nutrient levels in prawn farm effluent, a series of standardised short-term treatments were applied to various effluent types in a static 10,000-litre water body. Prawn pond effluents were characterised by watercolour and dominance of phytoplankton species. Three effluent types were tested, namely 1) particulate-rich effluent with little apparent phytoplankton, 2) green mircoalgal bloom predominately made up of single celled phytoplankton, and 3) brown microalgal bloom with higher prevalence of diatoms. The effluent types were similar (P>0.05) in non-volatile particulate material, and nitrate/nitrite but varied from each other in the following ways: 1) The particulate-rich effluents were lower (P<0.05) in volatile solids (compared to brown blooms), total Kjeldahl nitrogen, dissolved organic nitrogen, dissolved organic phosphorus and chlorophyll a (compared to both green and brown blooms). 2) The brown blooms were higher (P<0.05) in ammonia (compared to green blooms), total nitrogen and total phosphorus (compared to both green and particulate-rich effluent), but were lower (P<0.05) in inorganic phosphorus (compared to both green and particulate-rich effluent). 3) The green blooms were higher (P<0.05) in dissolved (both organic and inorganic) phosphorus (compared to both brown and particulate-rich effluents). Although the effluent types varied significantly in these aspects the effect of the Skim treatment was similar for all parameters measured except total phosphorus. Bloom type and Skim-treatment period significantly (P<0.05) affected total Kjeldahl phosphorus concentrations. For all effluent types there was a continuous significant reduction (P<0.05) in total Kjeldahl phosphorus during the initial 6-hour treatment period. Levels of total suspended solids and volatile suspended solids in all effluent types were significantly (P<0.05) reduced in the first 2 hours but not thereafter. Non-volatile suspended solids were also significantly (P<0.05) reduced in the first 2 hours (30 to 40 % reduction) and a further 40% reduction occurred in the particulate-rich effluent over the next 2 hours. Mean values for total ammonia, dissolved organic nitrogen, total Kjeldahl nitrogen, total nitrogen, chlorophyll a and dissolved organic or inorganic phosphorus levels were not significantly (P>0.05) affected by the Skim unit in any bloom type during the initial 6 hours of testing. Nevertheless, non-significant nitrogen reductions did occur. Foam production by the Skim unit varied with different blooms, resulting in different concentrate volumes and different end points for separate experiments. Concentrate volumes were generally high for the particulate-rich and green blooms (175 – 370 litres) and low for the brown blooms (25 – 80 litres). This was due to the low tendency of the brown bloom to produce foam. This generated higher nutrient concentrations in the associated condensed foam, but may have limited the treatment efficiency. The results suggest that in this application, the Skim unit did not remove micro-algae as effectively as was anticipated. However, it was effective at removing other suspended solids. Considering these attributes and the other uses of this machinery documented by the manufactures, the unit’s oxygenation mixing capacities coupled with inorganic solids removal may provide a suitable mechanism for construction of a continuously mixed bioreactor that utilises the filtration and profit making abilities of bivalves.

Utilisation of local ingredients in commercial pig feeds

The long-term competitiveness of the both the Vietnamese feed and pig production industries are constrained and under pressure whilst the industry is dependent on the use of imported feed ingredients in diets for animal production. These cost pressures are a result of import taxes, transport costs, currency fluctuations and feed supply limitations. By undertaking studies on available resources which are currently under-utilised and with potential as local feeds, we can prove their suitability for use as feedstuffs in pig diets and as replacements for imported feed ingredients. In undertaking this process we can lower feeding costs for pig production in Vietnam by the use of local feeds which are cheaper, generate new industries in Vietnam harvesting or processing these feeds and increase the incomes of Vietnamese workers who are involved in producing these by-products. Our project has shown that rubber seed, when processed correctly to lower the hydrogen cyanide content, is a safe and suitable protein meal feedstuff for use in pig diets with the potential to replace significant quantities of imported soybean and fishmeal in Vietnamese pig diets as long as diets are balanced for any amino acid shortfalls. Our peanut studies have shown that use of binders can help alleviate pig production problems with aflatoxin content in peanut meals. Further work is needed to characterise the fate of the bound aflatoxin to see if there is any meat residue risk. Cassava residue is a resultant by-product from starch extraction in both large and small cassava processing factories. Sub-samples from these two mill types were collected and evaluated for residue HCN. Analyses has shown that the processing and sun drying results in a product with relatively consistent low HCN content. Chemical analyses also reveal that significant residual starch also remains in this by-product. Digestibility studies and pig performance feeding studies have shown that cassava residue can be included in diets at 30% with no adverse effect, although the higher fibre content of this product means that strategically, cassava residue is more suitably used in finisher and sow diets. Research has examined the digestible energy content of a number of sunflower meal types available in Australia and identified major differences in their energy value based on processing, additionally, amino acid analysis has shown a significantly lower lysine content than previous reported. We also examined the digestible energy content of a number of Australian stylo forage legume harvest batches and identified the differences in their energy value based on age/harvest time of the forage legume. Analysis results of various stylo cuts showed that the early cut stylo has a higher starch content and lower fibre fraction content than observed in late and recut stylo which were allowed to grow longer. As a result the faecal digestible energy content was higher for the early cut stylo than for the subsequent cut stylo material which had been allowed to become woody. The results have shown that feeding of stylo meal does provide some nutritive value to the pig with increased energy and nitrogen supply, with a portion of the nitrogen presented which the pig is able to retain. Based on nutrient and fibre content stylo could have a useful role in sow feeding and satiety under non-stall housing situations. With increasing Vietnamese investment in rubber production seen with larger areas under plantations the amounts of rubber seed available for animal feeding will grow significantly over the next 15 years and the importance of the by-product ie rubber seed meal as a protein source in diets for Vietnamese pigs.