Nutritional constraints on displacement of Fusarium pseudograminearum from cereal straw by antagonists.

Displacement of the fungus Fusarium pseudograminearum from stubble by antagonists is a potential means of biocontrol of crown rot in cereals. The role of carbon and nitrogen nutrition in interactions between the pathogen and the antagonists Fusarium equiseti, Fusarium nygamai, Trichoderma harzianum and the non-antagonistic straw fungus Alternaria infectoria was investigated. Sole carbon source utilization patterns on Biolog plates were similar among the three Fusarium species, suggesting a possible role for competition. However, carbon niche overlap was unlikely to be important in antagonism by T. harzianum. Straw medium supplemented with sugars generally reduced the inhibitory effect of antagonists on growth of F. pseudograminearum in dual culture, indicating that availability of simple carbon sources does not limit antagonism. Adding nitrogen as urea, nitrate or ammonium to straw medium had little effect on antagonism by F. equiseti and F. nygamai, but ammonium addition removed the inhibitory effect of T. harzianum on growth of F. pseudograminearum. Displacement of F. pseudograminearum from straw by all fungi in a Petri dish assay was greater when urea or nitrate was used as a nitrogen source than with ammonium. All forms of nitrogen significantly increased displacement of F. pseudograminearum from straw under simulated field conditions when straws were either inoculated with T. harzianum or exposed to resident soil microbes. However, in 2 out of 3 experiments urea and nitrate were more effective than ammonium. The results suggest that availability of nitrogen, but not carbon, is limiting the activities of antagonists of F. pseudograminearum in straw, and the way nitrogen is applied can influence the rate of displacement and mortality of the pathogen in host residues.

An evaluation of electrofishing as a control measure for an invasive tilapia (Oreochromis mossambicus) population in northern Australia.

Combating the spread of invasive fish is problematic, with eradication rarely possible and control options varying enormously in their effectiveness. In two small impoundments in north-eastern Australia, an electrofishing removal program was conducted to control an invasive tilapia population. We hypothesised that electrofishing would reduce the population density of Oreochromis mossambicus (Mozambique tilapia), to limit the risk of downstream spread into areas of high conservation value. We sampled by electrofishing monthly for 33 months. Over this period, there was an 87% decline in catch per unit effort (CPUE) of mature fish, coupled with a corresponding increase of 366% in the number of juveniles, suggesting a density-dependent response in the stock-recruitment relationship for the population. Temperature was inversely related to CPUE (r=0.43, lag=10 days), implying greater electrofishing efficiency in cooler months. The reduction in breeding stock is likely to reduce the risk of spread and render the population vulnerable to other control measures such as netting and/or biological control. Importantly, the current study suggests routine electrofishing may be a useful control tool for invasive fish in small impoundments when the use of more destructive techniques, such as piscicides, is untenable.

Commercial application of polychaete sand filters for wastewater remediation and broodstock feeds

Summary Prototype sand-worm filtration beds were constructed at two prawn farms and one fish farm to assess and demonstrate their polychaete (marine worm) production and wastewater remediation capacities at semi-commercial scale. Wastewater treatment properties were monitored and worms produced were assessed and either sold for bait or used by the farms’ hatcheries as broodstock (prawn or fish breeder) feed. More than 34 megalitres of prawn- and fish-pond water was beneficially treated in the 116-319-d trial. The design of the polychaete-assisted sand filters (PASFs) constructed at each farm affected their water handling rates, which on average ranged from 315 to 1000 L m-2 d-1 at the three farms. A low profile design incorporating shallow bunded ponds made from polyethylene liner and timber stakes provided the easiest method of construction. This simple design applied at broad scale facilitated the highest quantities of treated water and the greatest worm production. Designs with higher sides increased the head pressure above the sand bed surface, thus increasing the amount of water that could be treated each day. Most water qualities were affected in a similar way to that demonstrated in the previous tank trials: dissolved oxygen, pH, total suspended solids and chlorophyll a levels were all consistently significantly lowered as pond water percolated through the sand bed, and dissolved forms of nitrogen and phosphorus were marginally increased on several occasions. However, unlike the previous smaller-scale tank trials, total nitrogen (TN) and total phosphorus (TP) levels were both significantly lowered by these larger-scale PASFs. The reasons for this are still unclear and require further research. Maximum TN and TP removals detected in the trial were 48.8% and 67.5%, respectively, and average removals (in unfed beds) at the three farms ranged from 20.0 to 27.7% for TN and from 22.8 to 40.8% for TP. Collectively, these results demonstrate the best suspended solids, chlorophyll and macronutrient removal capacities so far reported for any mariculture wastewater treatment methodology to date. Supplemental feeding of PASFs with fish meal was also investigated at one farm as a potential means of increasing their polychaete biomass production. Whilst fed beds produced higher biomass (152 ± 35 g m-2) compared with unfed beds (89 ± 17 g m-2) after 3.7 months of operation, the low number of replicates (2) prevented statistically significant differences from being demonstrated for either growth or survival. At harvest several months later, worm biomass production was estimated to be similar to, or in slight excess of, previously reported production levels (300-400 g m-2). Several qualities of filtered water appear to have been affected by supplemental feeding: it appeared to marginally lower dissolved oxygen and pH levels, and increased the TN and TP levels though not so much to eliminate significant beneficial water treatment effects. Periodic sampling during an artificial-tide demonstrated the tendency for treated-water quality changes during the first hour of filtration. Total nitrogen and ammonia peaked early in the tidal flow and then fell to more stable levels for the remainder of the filtration period. Other dissolved nutrients also showed signs of this sand-bed-flushing pattern, and dissolved oxygen tended to climb during the first hour and become more stable thereafter. These patterns suggest that the routine sampling of treated water undertaken at mid-inflow during the majority of the wider study would likely have overestimated the levels of TN and dissolved nutrients discharged from the beds, and hence underestimated the PASFs treatment efficacies in this regard. Analyses of polychaete biomass collected from each bed in the study revealed that the worms were free from contamination with the main prawn viruses that would create concerns for their feeding to commercial prawn broodstock in Australia. Their documented proximal and nutritional contents also provide a guide for hatchery operators when using live or frozen stock. Their dry matter content ranged from 18.3 to 22.3%, ash ranged from 10.2 to 14.0%, gross energy from 20.2 to 21.5 MJ kg-1, and fat from 5.0 to 9.2%. Their cholesterol levels ranged from 0.86 to 1.03% of dry matter, whilst total phospholipids range from 0.41 to 0.72%. Thirty-one different fatty acids were present at detectable (≥0.005% of dry matter) levels in the sampled worm biomass. Palmitic acid was by far the most prevalent fatty acid detected (1.21 ± 0.18%), followed by eicosapentaenoic (EPA) (0.48 ± 0.03%), stearic (0.46 ± 0.04%), vaccenic (0.38 ± 0.05%), adrenic (0.35 ± 0.02%), docosadienoic (0.28 ± 0.02%), arachidonic (AA) (0.22 ± 0.01%), palmitoleic (0.20 ± 0.04%) and 23 other fatty acids with average contents of less than 0.2% of dry matter. Supplemental feeding with fish meal at one farm appeared to increase the docosahexaenoic acid (DHA) content of the worms considerably, and modify the average AA : EPA : DHA from 1.0 : 2.7 : 0.3 to 1.0 : 2.0 : 1.1. Consistent with previous results, the three most heavily represented amino acids in the dry matter of sampled worms were glutamic acid (8.5 ± 0.2%), aspartic acid (5.5 ± 0.1%) and glycine (4.9 ± 0.5%). These biomass content results suggest that worms produced in PASF systems are well suited to feeding to prawn and fish broodstock, and provide further strong evidence of the potential to modify their contents for specific nutritional uses. The falling wild-fishery production of marine bloodworms in Queensland is typical of diminishing polychaete resources world-wide and demonstrates the need to develop sustainable production methods here and overseas. PASF systems offer the dual benefits of wastewater treatment for environmental management and increased productivity through a valuable secondary crop grown exclusively on waste nutrients.

Polychaete-assisted sand filters – prawn farm wastewater remediation trial

Medium bedding sand which is commonly available in coastal sedimentary deposits, and a marine polychaete-worm species from Moreton Bay recently classified as Perinereis helleri (Nereididae), were deployed in a simple low-maintenance sand filter design that potentially has application at large scale. Previous work had shown that this physical and biological combination can provide a new option for saline wastewater treatment, since the worms help to prevent sand filter blocking with organic debris and offer a profitable by-product. To test the application of this new concept in a commercial environment, six 1.84 m2 Polychaete-assisted sand filters were experimentally tested for their ability to treat wastewater from a semi-intensive prawn culture pond. Polychaetes produced exclusively on the waste nutrients that collected in these gravity-driven sand filters were assessed for their production levels and nutritional contents. Water parameters studied included temperature, salinity, pH, dissolved oxygen (DO), oxidation/ reduction potential (redox), suspended solids, chlorophyll a, biological oxygen demand (BOD), and common forms of nitrogen and phosphorus. Pond water which had percolated through the sand bed had significantly lower pH, DO and redox levels compared with inflow water. Suspended solids and chlorophyll a levels were consistently more than halved by the process. Reductions in BOD appeared dependant on regular subsurface flows. Only marginal reductions in total nitrogen and phosphorus were documented, but their forms were altered in a potentially useful way: dissolved forms (ammonia and orthophosphate) were generated by the process, and this remineralisation also seemed to be accentuated by intermittent flow patterns. Flow rates of approximately 1,500 L m-2 d-1 were achieved suggesting that a 1 ha polychaete bed of this nature could similarly treat the discharge from a 10 ha semi-intensive prawn farm. Sixteen weeks after stocking sand beds with one-month-old P. helleri, over 3.6 kg of polychaete biomass (wet weight) was recovered from the trial. Production on a sand bed area basis was 328 g m-2. Similar (P>0.05) overall biomass production was found for the two stocking densities tested (2000 and 6000 m-2; n = 3), but survival was lower and more worms were graded as small (<0.6 g) when produced at the higher density (28.2 ± 1.5 % and approx. 88 %, respectively) compared with the lower density (46.8 ± 4.4 % and approx. 76 %, respectively). When considered on a weight for weight basis, about half of the worm biomass produced was generally suitable for use as bait. The nutritional contents of the worms harvested were analysed for different stocking densities and graded sizes. These factors did not significantly affect their percentages of dry matter (DM) (18.23 ± 0.57 %), ash (19.77 ± 0.80 % of DM) or gross energy 19.39 ± 0.29 MJ kg-1 DM) (n = 12). Although stocking density did not affect the worms’ nitrogen and phosphorus contents, small worms had a higher mean proportion of nitrogen and phosphorus (10.57 ± 0.17 % and 0.70 ± 0.01 % of DM, respectively) than large worms (9.99 ± 0.12 % and 0.65 ± 0.01 % of DM, respectively) (n = 6). More lipid was present in large worms grown at the medium density (11.20 ± 0.19 %) compared with the high density (9.50 ± 0.31 %) and less was generally found in small worms (7.1-7.6 % of DM). Mean cholesterol and total phospholipid levels were 5.24 ± 0.15 mg g-1 and 13.66 ± 2.15 mg g-1 DM, respectively (n = 12). Of the specific phospholipids tested, phosphatidyl-serine or sphingomyelin were below detection limits (<0.05 mg g-1), whilst mean levels of phosphatidyl-ethanolamine, phosphatidyl-inositol, phosphatidyl-choline and lysophosphatidyl-choline were 6.89 ± 1.09, 0.89 ± 0.26, 4.04 ± 1.17 and 1.84 ± 0.37 mg g-1, respectively (n = 12). Culture density generally had a more pronounced effect on phospholipid contents than did size of worms. By contrast, worm size had a more pronounced effect on total fatty acid contents, with large worms containing significantly higher (P<0.001) levels on a DM basis (46.88 ± 2.46 mg g-1) than smaller worms (27.76 ± 1.28 mg g-1). A very broad range of fatty acids were detected with palmitic acid being the most heavily represented class (up to 14.23 ± 0.49 mg g-1 DM or 27.28 ± 0.22 % of total fatty acids). Other heavily represented classes included stearic acid (7.4-8.8 %), vaccenic acid (6.8-7.8 %), arachidonic acid (3.5-4.4 %), eicosapentaenoic acid (9.9-13.8 %) and docosenoic acid (5.7-7.0 %). Stocking density did not affect (P>0.05) the levels of amino acids present in polychaete DM, but there was generally less of each amino acid tested on a weight per weight basis in large worms than in small worms. This difference was significant (P<0.05) for the most heavily represented classes being glutamic acid (73-77 mg g-1), aspartic acid (50-54 mg g-1), and glycine (46-53 mg g-1). These results demonstrate how this polychaete species can be planted and sorted at harvest according to various strategies aimed at providing biomass with specific physical and nutritional qualities for different uses.

Using molasses to control inorganic nitrogen and pH in aquaculture ponds

This article describes research designed to determine the molasses addition rates that can control ammonia and pH in eutrophic aquaculture ponds.

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.

Relative host plant species use by the lantana biological control agent Aconophora compressa (Membracidae) across its native and introduced ranges

Aconophora compressa Walker (Hemiptera: Membracidae) was released in 1995 against the weed lantana in Australia, and is now found on multiple host plant species. The intensity and regularity at which A. compressa uses different host species was quantified in its introduced Australian range and also its native Mexican range. In Australia, host plants fell into three statistically defined categories, as indicated by the relative rates and intensities at which they were used in the field. Fiddlewood (Citharexylum spinosum L.: Verbenaceae) was used much more regularly and at higher densities than any other host sampled, and alone made up the first group. The second group, lantana (Lantana camara L.: Verbenaceae; pink variety) and geisha girl (Duranta erecta L.: Verbenaceae), were used less regularly and at much lower densities than fiddlewood. The third group, Sheena’s gold (another variety of D. erecta), jacaranda (Jacaranda mimosifolia D. Don: Bignoniaceae) and myoporum (Myoporum acuminatum R. Br.: Myoporaceae), were used infrequently and at even lower densities. In Mexico, the insect was found at relatively low densities on all hosts relative to those in Australia. Densities were highest on L. urticifolia, D. erecta and Tecoma stans (L.) Juss. ex Kunth (Bignoniaceae), which were used at similar rates to one another. It was found also on a few other verbenaceous and non-verbenaceous host species but at even lower densities. The relative rate at which Citharexylum spp. and L. urticifolia were used could not be assessed in Mexico because A. compressa was found on only one plant of each species in areas where these host species co-occurred. The low rate at which A. compressa occurred on fiddlewood in Mexico is likely to be an artefact of the short-term nature of the surveys or differences in the suites of Citharexylum and Lantana species available there. These results provide further incentive to insist on structured and quantified surveys of non-target host use in the native range of potential biological control agents prior to host testing studies in quarantine.

Assessing the biological control potential of an adventitiously-established "pest", Scirtothrips aurantii (Faure), on a weed, Bryophyllum delagoense (Eckl. & Zeyh.), in Queensland

South African citrus thrips (Scirtothrips aurantii) established adventitiously in Australia. Although it is a major horticultural pest in Africa, it is now advocated as a possible biological control agent against Bryophyllum delagoense Eckl. & Zeyh. (Crassulaceae). To evaluate the biocontrol potential of S. aurantii a two year field study was conducted on the western Darling Downs of southern Queensland. Imidacloprid insecticide was applied to two quadrats at each of 18 field sites to assess, in the absence of S. aurantii, the persistence of individual plants and to quantify propagule production and recruitment by this declared weed. A third quadrat was left, as a control, to be infested naturally by S. aurantii. When released from herbivory by thrips in the field, plants grew significantly more, flowered more, and were significantly more fecund than plants in the quadrats with S. aurantii. Increases in growth and fecundity translated into significantly increased plant numbers but not increased recruitment. Recruitment even declined in experimental quadrats, through the indirect effects of releasing plants from herbivory. Field sampling also revealed that S. aurantii may be sensitive to seasonal climatic fluctuations. These and other local climatic influences may limit the biological control potential of the insect.

Trialling Biological Agents for the Management of Lesser Mealworm in Australian Broiler Houses

For approximately three decades the Australian broiler industry has relied heavily on the use of insecticides as its key tool for management of darkling beetle or lesser mealworm, Alphitobius diaperinus [Panzer] in broiler houses. The use of these chemicals over this period has been largely unchecked which has resulted in the development of strong insecticide resistance in many beetle populations from broiler farms. Although we are in a period now with an improved knowledge of managing resistance and the availability of new more effective insecticides that are currently marketed, the industry still requires more pest management options in order to inhibit development of resistance and reduce overall chemical use. In response to this need, ‘natural’ agents such as entomopathogenic nematodes and fungi were proposed as potential agents for managing darkling beetle populations in Australian broiler houses. Since 2007 laboratory and field studies have been undertaken to assess these agents. This report outlines these studies and discusses potential benefits to the Chicken Meat industry resulting from this research.

Biological Control of Eucalypt Pests Overseas and Australia

This project aims to address the growing need for a coordinated approach to research into the biological control of Australian eucalypt insect pests by scoping the formation of a Centre in Australia which would (a) coordinate the evaluation and provision of biological control agents (initially to South Africa and Brazil, but in future years more widely), (b) research the role natural enemies play in pest population regulation in Australian eucalypt plantations and how this may be enhanced as a management tool, and (c) form a network focussed on forest biosecurity with an emphasis on eucalypt pests and pathogens.

Biological suppression of root-lesion nematodes in grain-growing soils

Root-lesion nematodes (RLNs) are found on 75% of grain farms in southern Queensland (QLD) and northern New South Wales (NSW) and are significant pests. This project confirmed that biological suppression of RLNs occurs in soils, examined what organisms are involved and how growers might enhance suppressiveness of soils. Field trials, and glasshouse and laboratory bioassays of soils from fields with contrasting management practices, showed suppressiveness is favoured with less tillage, more stubble and continuous intensive cropping, particularly in the top 15cm of soil. Through extensive surveys key organisms, Pasteuria bacteria, nematode-trapping fungi and predatory nematodes were isolated and identified as being present.

Nitrogen requirements of Burdekin soils to inform Reef Policy legislation

Reef Project 20: Nitrogen fertiliser requirements of representative soils of the Burdekin (BRIA and Delta), and peaty soils of the Wet Tropics to inform the ReefWise farming Nutrient Calculator.

Optimising nitrogen fixation of grain and forage legumes in the northern region

This project encompasses laboratory, glasshouse and field research to improve N fixation in grain and forage legumes in the northern region and assess compatability of rhizobial strains with current and new legume varieties.

Life history and host range of Alagoasa extrema (Coleoptera: Chrysomelidae: Alticinae), a potential biological control agent of Lantana spp. (Verbenaceae) in Australia

The life history and host range of the lantana beetle, Alagoasa extrema, a potential biocontrol agent for Lantana spp. were investigated in a quarantine unit at the Alan Fletcher Research Station, Brisbane, Australia. Adults feed on leaves and females lay batches of about 17 eggs on the soil surface around the stems of plants. The eggs take 16 days to hatch and newly emerged larvae move up the stem to feed on young leaves. Larvae feed for about 23 days and there are three instars. There is a prepupal non-feeding stage that lasts about 12 days and the pupal stage, which occurs in a cocoon in the soil, lasts 16 days. Teneral adults remain in the cocoon for 3 days to harden prior to emergence. Males live for about 151 days while females live for about 127 days. The pre-oviposition period is 19 days. In no-choice larval feeding trials, nine plant species, representing three families, supported development to adult. Three species, Aloysia triphylla, Citharexylum spinosum and Pandorea pandorana were able to support at least two successive generations. These results confirm those reported in South Africa and suggest that A. extrema is not sufficiently specific for release in Australia. Furthermore, it is not recommended for release in any other country which is considering biological control of lantana.

Using Trichogramma Westwood (Hymenoptera: Trichogrammatidae) for insect pest biological control in cotton crops: an Australian perspective.

Trichogramma Westwood egg parasitoids alone generally fail to suppress heliothine pests when released in established cotton-growing regions. Factors hindering their success include indiscriminate use of detrimental insecticides, compensation for minimal pest larval hatch due to their activity via reduced larval cannibalism or mortality in general, singly laid heliothine eggs avoiding detection and asynchronous development benefiting host over parasitoid. Yet, despite these limitations, relatively large Trichogramma pretiosum Riley populations pervade and effectively suppress Helicoverpa (Hardwick) pests in Australian Bt (Bacillus thuringiensis Berliner)-transgenic cotton, Gossypium hirsutum L., crops, especially in the Ord River Irrigation Area (ORIA) of tropical northern Australia, where their impact on the potentially resistant pest species, Helicoverpa armigera (Hubner), is considered integral to the local insecticide resistance management (IRM) strategy for continued, sustainable Bt-transgenic cotton production. When devoid of conventional insecticides, relatively warm and stable conditions of the early dry season in winter grown ORIA Bt-transgenic cotton crops are conducive to Trichogramma proliferation and biological control appears effective. Further, there is considerable scope to improve Trichogramma's biological control potential, in both the ORIA and established cotton-growing regions, via habitat manipulation. It is proposed that Trichogramma may prove equally effective in developing agricultural regions of monsoonal northern Australia, and that environmental constraints on Trichogramma survival, and those of other natural enemies, require due consideration prior to their successful application in biological control programs.