Tertiary Treatment of Ayr Municipal Wastewater using Bioremediation: A Pilot-Scale Study. A Report Prepared for the Burdekin Shire Council and the Burdekin Rangeland Reef Initiative

This joint DPI/Burdekin Shire Council project assessed the efficacy of a pilot-scale biological remediation system to recover Nitrogen (N) and Phosphorous (P) nutrients from secondary treated municipal wastewater at the Ayr Sewage Treatment Plant. Additionally, this study considered potential commercial uses for by-products from the treatment system. Knowledge gained from this study can provide directions for implementing a larger-scale final effluent treatment protocol on site at the Ayr plant. Trials were conducted over 10 months and assessed nutrient removal from duckweed-based treatments and an algae/fish treatment – both as sequential and as stand-alone treatment systems. A 42.3% reduction in Total N was found through the sequential treatment system (duckweed followed by algae/fish treatment) after 6.6 days Effluent Retention Time (E.R.T.). However, duckweed treatment was responsible for the majority of this nutrient recovery (7.8 times more effective than algae/fish treatment). Likewise, Total P reduction (15.75% reduction after 6.6 days E.R.T.) was twice as great in the duckweed treatment. A phytoplankton bloom, which developed in the algae/fish tanks, reduced nutrient recovery in this treatment. A second trial tested whether the addition of fish enhanced duckweed treatment by evaluating systems with and without fish. After four weeks operation, low DO under the duckweed blanket caused fish mortalities. Decomposition of these fish led to an additional organic load and this was reflected in a breakdown of nitrogen species that showed an increase in organic nitrogen. However, the Dissolved Inorganic Nitrogen (DIN: ammonia, nitrite and nitrate) removal was similar between treatments with and without fish (57% and 59% DIN removal from incoming, respectively). Overall, three effluent residence times were evaluated using duckweed-based treatments; i.e. 3.5 days, 5.5 days and 10.4 days. Total N removal was 37.5%, 55.7% and 70.3%, respectively. The 10.4-day E.R.T. trial, however, was evaluated by sequential nutrient removal through the duckweed-minus-fish treatment followed by the duckweed-plus-fish treatment. Therefore, the 70.3% Total N removal was lower than could have been achieved at this retention time due to the abovementioned fish mortalities. Phosphorous removal from duckweed treatments was greatest after 10.4-days E.R.T. (13.6%). Plant uptake was considered the most important mechanism for this P removal since there was no clay substrate in the plastic tanks that could have contributed to P absorption as part of the natural phosphorous cycle. Duckweed inhibited phytoplankton production (therefore reducing T.S.S) and maintained pH close to neutral. DO beneath the duckweed blanket fell to below 1ppm; however, this did not limit plant production. If fish are to be used as part of the duckweed treatment, air-uplifts can be installed that maintain DO levels without disturbing surface waters. Duckweed grown in the treatments doubled its biomass on average every 5.7 days. On a per-surface area basis, 1.23kg/m2 was harvested weekly. Moisture content of duckweed was 92%, equating to a total dry weight harvest of 0.098kg/m2/week. Nutrient analysis of dried duckweed gave an N content of 6.67% and a P content of 1.27%. According to semi-quantitative analyses, harvested duckweed contained no residual elements from the effluent stream that were greater than ANZECC toxicant guidelines proposed for aquaculture. In addition, jade perch, a local aquaculture species, actively consumed and gained weight on harvested duckweed, suggesting potential for large-scale fish production using by-products from the effluent treatment process. This suggests that a duckweed-based system may be one viable option for tertiary treatment of Ayr municipal wastewater. The tertiary detention lagoon proposed by the Burdekin Shire Council, consisting of six bays approximately 290 x 35 metres (x 1.5 metres deep), would be suitable for duckweed culture with minor modification to facilitate the efficient distribution of duckweed plants across the entire available growing surface (such as floating containment grids). The effluent residence time resulting from this proposed configuration (~30 days) should be adequate to recover most effluent nutrients (certainly N) based on the current trial. Duckweed harvest techniques on this scale, however, need to be further investigated. Based on duckweed production in the current trial (1.23kg/m2/week), a weekly harvest of approximately 75 000kg (wet weight) could be expected from the proposed lagoon configuration under full duckweed production. A benefit of the proposed multi-bay lagoon is that full lagoon production of duckweed may not be needed to restore effluent to a desirable standard under the present nutrient load, and duckweed treatment may be restricted to certain bays. Restored effluent could be released without risk of contaminating the receiving waterway with duckweed by evacuating water through an internal standpipe located mid-way in the water column.

Effects of secondary crops on bacterial growth and nitrogen removal in shrimp farm effluent treatment systems

Secondary crops provide a means of assimilating some effluent nitrogen from eutrophic shrimp farm settlement ponds. However, a more important role may be their stimulation of beneficial bacterial nitrogen removal processes. In this study, bacterial biomass, growth and nitrogen removal capacity were quantified in shrimp farm effluent treatment systems containing vertical artificial substrates and either the banana shrimp Penaeus merguiensis (de Man) or the grey mullet, Mugil cephalus L. Banana shrimp were found to actively graze biofilm on the artificial substrates and significantly reduced bacterial biomass relative to a control (24.5 ± 5.6mgCm−2 and 39.2 ± 8.7mgCm−2, respectively). Bacterial volumetric growth rates, however, were significantly increased in the presence of the shrimp relative to the control 45.2±11.3mgCm−2 per day and 22.0±4.3mgCm−2 per day, respectively). Specific growth rate, or growth rate per cell, of bacteria was therefore appreciably stimulated by the banana shrimp. Nitrate assimilation was found to be significantly higher on grazed substrate biofilm relative to the control (223±54 mgNm−2 per day and 126±36 mg Nm−2 per day, respectively), suggesting that increased bacterial growth rate does relate to enhanced nitrogen uptake. Regulated banana shrimp feeding activity therefore can increase the rate of newbacterial biomass production and also the capacity for bacterial effluent nitrogen assimilation. Mullet had a negligible influence on the biofilm associated with the artificial substrate but reduced sediment bacterial biomass (224 ± 92 mgCm−2) relative to undisturbed sediment (650 ± 254 mgCm−2). Net, or volumetric bacterial growth in the sediment was similar in treatments with and without mullet, suggesting that the growth rate per cell of bacteria in grazed sediments was enhanced. Similar rates of dissolved nitrogen mineralisation werefound in sediments with and without mullet but nitrificationwas reduced. Presence of mullet increased water column suspended solids concentrations, water column bacterial growth and dissolved nutrient uptake. This study has shown that secondary crops, particularly banana shrimp, can play a stimulatory role in the bacterial processing of effluent nitrogen in eutrophic shrimp effluent treatment systems.

Treatment of shrimp farm effluent with omnivorous finfish and artificial substrates

Long-term environmental sustainability and community acceptance of the shrimp farming industry in Australia requires on-going development of efficient cost-effective effluent treatment options. In this study, we aimed to evaluate the effectiveness of a shrimp farm treatment system containing finfish and vertical artificial substrates (VAS). This was achieved by (1) quantifying the individual and collective effects of grey mullet (Mugil cephalus L.) and VASs on water and sediment quality, and (2) comparing the retention of N in treatment systems with and without the presence of finfish (M. cephalus and the siganid Siganus nebulosus (Quoy & Gaimard)), where light was selectively removed. Artificial substrates were found to significantly improve the settlement of particulate material, regardless of the presence of finfish. Mullet actively resuspended settled solids and reduced the production of nitrate when artificial substrates were absent. However, appreciable nitrification was observed when mullet were present together with artificial substrates. The total quantity of N retained by the mullet was found to be 1.8– 2.4% of the incoming pond effluent N. It was estimated that only 21% of the pond effluent N was available for mullet consumption. When S. nebulosus was added, total finfish N retention increased from 1.8% to 3.9%, N retention by mullet also improved (78±16 to 132±21-mg N day−1 before and after siganid addition respectively). Presence of filamentous macroalgae (Enteromorpha spp.) was found to improve the removal of N from pond effluent relative to treatments where light was excluded. Denitrification was also a significant sink for N (up to 24% N removed). Despite the absence of algal productivity and greater availability of nitrate, denitrification was not higher in treatments where light was excluded. Mullet were found to have no effect on the rates of denitrification but significantly reduced macroalgal growth on the surface of the water. When mullet were absent, excessive macroalgal growth led to reduced dissolved oxygen concentrations and nitrification. This study concludes that the culture of mullet alone in shrimp farm effluent treatment systems does not result in significant retention of N but can contribute to the control of macroalgal biomass. To improve N retention and removal, further work should focus on polyculturing a range of species and also on improving denitrification.

Phenotypic and genotypic variation within populations of kikuyu (Pennisetum clandestinum) in Australia.

Experiments at 2 sites in subtropical eastern Australia investigated the variation in agronomic attributes, quality and genetic structure existing within: naturally-occurring populations of kikuyu ( Pennisetum clandestinum) from within Australia; selections produced from the treatment of Whittet seed with mutagenic chemicals; and available cultivars. Runners were collected from coastal areas extending from Western Australia to the Atherton Tableland in north Queensland. One experiment evaluated 10 mutagenic selections and 4 cultivars in a lattice design and the other evaluated 12 ecotypes and 3 cultivars in a randomised block design. The experimental unit was single plants, which were sown on a 1.5 m grid into a weed-free seed-bed (Mutdapilly) or a killed kikuyu stand (Wollongbar), both of which were kept clear of weeds and other kikuyu plants for the duration of the experiments. Foliage height, forage production and runner yield were assessed. Leaf material was analysed for concentrations of crude protein (CP), acid detergent fibre (ADF) and neutral detergent fibre (NDF) and for in vitro dry matter digestibility (IVDDM) in autumn, winter and spring. DNA was extracted from each plant in the ecotype comparison and subjected to a modified DAF (DNA amplification fingerprinting) analysis to determine the level of genetic relatedness. In the first experiment, none of the mutagenic lines derived from Whittet yielded significantly more or was more digestible than commercial Whittet material, although some selections were superior to the other commercial kikuyu cultivars, Noonan and Crofts, and 'common' kikuyu. However, there were significant differences in plant height and runner expansion. In the second experiment, significant differences in plant height, foliage yield, runner development, and leaf CP, ADF, NDF and IVDDM concentrations were demonstrated between the ecotypes, mutagenic selections and cultivars. There was a 4- to 6-fold difference in plant yield and a 6- to 10-fold difference in runner production between the ecotypes at the 2 sites. Quality of the leaf ranged from 200 to 270 g/kg (CP), from 700 to 770 g/kg (IVDDM), from 170 to 250 g/kg (ADF) and from 470 to 550 g/kg (NDF). Improvements in quality and agronomic attributes were not mutually exclusive. Genetic fingerprint analysis of the kikuyu lines indicated that they formed 2 broad groupings. Most of the regional ecotypes were grouped with 'common' kikuyu as represented by the material collected from Wollongbar, and the Beechmont, Atherton Tableland and Gympie ecotypes were grouped with the registered cultivars Whittet, Noonan and Crofts. Two lines produced by mutagenesis from Whittet remained closely linked to Whittet. These results suggest that there was variation between populations of kikuyu in yield, quality and genetic diversity but that mutagenesis by treating seed with sodium azide and diethylene sulphide did not achieve a significant change in the digestibility of leaf over cv. Whittet.

Managing the grass-legume balance in Stylosanthes scabra cv. Seca pastures in central Queensland

The impacts of 4 grazing strategies (year-long grazing, summer grazing, winter grazing and winter grazing plus spring burning) on the grass:legume balance were studied between 2000 and 2006 in a pasture oversown with Stylosanthes scabra cv. Seca (Seca stylo) in central Queensland. Seasonal rainfall throughout the study was generally below average. Total pasture yields in autumn were higher in the 2 winter grazing than the 2 summer grazing treatments, largely reflecting the sampling time relative to when grazing occurred. There were few differences in Seca composition in autumn, although there was a clear trend for Seca composition to be reduced by winter grazing plus burning. Both the frequency of occurrence and plant density of Seca were higher under the 2 summer grazing treatments and there was also a trend for the density of juvenile plants (<5 cm height) to be higher in the 2 summer grazing treatments. Seca soil seed banks were generally low and were reduced in the winter grazing plus burning treatment in spring 2002. The frequency of the palatable perennial grass Pennisetum ciliaris (Biloela buffel grass) was reduced while that of the 'increaser' species Bothriochloa pertusa (Indian couch grass) and Stachytarpheta jamaicensis (snake weed) increased in the 2 summer grazing treatments compared with the 2 winter grazing treatments. Burning in spring increased soil loss in treatments grazed in winter. Differences in Seca frequency and density but not composition were explained by the 2 summer grazing treatments promoting 'gaps' in the pasture which were then colonised by Seca plants and other 'increaser' species. It was reasoned that, with time, mature Seca plants in the 2 winter grazing treatments would die so that Seca composition would eventually become higher under summer grazing regimes than under winter grazing. It was concluded that limiting grazing to particular seasons can alter legume:grass balance and that a time-frame of 5-8 years with average to good rainfall would be necessary to achieve large shifts in composition.

Haemophilus parasuis treatment

The aim of the project is to prove that live vaccination on the farm will protect the piglets from heterologous challenge with H. parasuis. The steps to achieve the aim of the project are to find a dose rate on the farm which guarantees colonisation of the vaccine strain and is safe On farm vaccinated and unvaccinated pigs are then shifted to CAAS at three weeks of age and challenged with a heterologous strains. The method is then applied on a large piggery for a period of nearly 12 months. We will also develop a freeze drying method and a technical manual of procedures to identify serovars prevailing on pig farms and which serovar to include into the vaccine.

DAQ339A Pest and disease identification and management in herbs

The major objective is to produce an educational tool for growers and research/extension personnel to allow accurate identification of a range of pests and diseases encountered in herbs. To a lessor extent develop both a mechanism to manage beneficial insects in field crops pre-harvest and to identify some common seed borne diseases in herbs.

Developing and communicating strategies for controlling virus diseases in vegetable cucurbit crops

Virus diseases cause serious yield and quality losses in field grown cucurbit crops worldwide. In Australia, the main viruses of cucurbits are Papaya ringspot virus (PRSV), Squash mosaic virus (SqMV), Watermelon mosaic virus (WMV) and Zucchini yellow mosaic virus (ZYMV). Plants infected early have severely distorted fruit. High infection incidences, of ZYMV and PRSV in crops cause losses of marketable fruit of up to 100% and infected crops are often abandoned. Two new alternative hosts of ZYMV were identified, the native cucurbit Cucumis maderaspatanus and wild legume Rhyncosia minima. No new alternative hosts of PRSV, SqMV or WMV were found in Western Australia or Queensland. Seed transmission of ZYMV (0.7%) was found in seedlings grown from ZYMV-infected fruit of zucchini but not of pumpkin. None was detected with PRSV or SqMV in zucchini or pumpkin seedlings, respectively. ZYMV spread to pumpkins by aphids was greater downwind than upwind of a virus source. Delaying sowing by 2 weeks decreased ZYMV spread. Millet non-host barriers between pumpkin plantings slowed ZYMV infection. Host resistance gene (zym) in cucumber cultivars was effective against ZYMV. Pumpkin cultivars with resistance gene (Zym) became infected under high virus pressure but leaf symptoms were milder and infected plants higher yielding with more market-acceptable fruit than those without Zym. Most zucchini cultivars with Zym developed severe leaf and fruit symptoms. ZYMV, PRSV, WMV and SqMV spread readily from infected to healthy cucurbit plants by direct leaf contact. ZYMV survives and remains infective on diverse surfaces for up to 6 hours but can be inactivated by some disinfectants. Phylogenetic analysis indicates at least three separate introductions of ZYMV into Australia, with new introductions rarely occurring. ZYMV isolates clustered into three groups according to collection location i) Kununurra, ii) Northern Territory and iii) Carnarvon, Qld and Vic. A multiplex Real-Time PCR was developed which distinguished between the three groups of Australian isolates. Integrated disease management (IDM) strategies for virus diseases of vegetable cucurbit crops grown in the field were improved incorporating the new information gathered. These strategies are aimed at causing using minimal extra expense, labour demands and disruption to normal practices.

Vapour heat treatment of honey gold mango

Vapour heat treatment of honey gold mango for access to the Japanese market.

Alternative fruit fly treatment for interstate market access for strawberries

This project aims to provide further data to support the winter window option for interstate marker access for strawberries from south east Queensland.

UWA00125 Weed Seed Wizard

The aim of this project is to further develop, improve and validate the Weed Seed Wizard, the weed seedbank management tool that was developed in the Weed CRC and to encourage its adoption by researchers, growers and consultants. The focus of the first phase of the project (Phase One) will be on calibrating and validating the model against experimental biological data and trial and paddock monitoring results, collected from published scientific literature, previous studies and ongoing trials and experiments.

Innovative management of herbicide resistant weeds - seed banks and soil biology

The project will evaluate seed bank depletion of key northern herbicide resistant weeds under different environments, cropping systems, crop agronomies and non-chemical control tactics. The project will also evaluate soil biology and seed bank relationships to explain differences in seed bank persistence.

Improving management of summer weeds in dryland cropping systems with Cotton

In 2001 a scoping study (phase I) was commissioned to determine and prioritise the weed issues of cropping systems with dryland cotton. The main findings were that the weed flora was diverse, cropping systems complex, and weeds had a major financial and economical impact. Phase II 'Best weed management strategies for dryland cropping systems with cotton' focused on improved management of the key weeds, bladder ketmia, sowthistle, fleabane, barnyard grass and liverseed grass.In Phase III 'Improving management of summer weeds in dryland cropping systems with cotton', more information on the seed-bank dynamics of key weeds was gained in six pot and field studies. The studies found that these characteristics differed between species, and even climate in the case of bladder ketmia. Species such as sowthistle, fleabane and barnyard grass emerged predominately from the surface soil. Sweet summer grass was also in this category but also had a significant proportion emerging from 5 cm depth. Bladder ketmia in central Queensland emerged mainly from the top 2 cm, whereas in southern Queensland it emerged mainly from 5 cm. Liverseed grass had its highest emergence from 5 cm below the surface. In all cases the persistence of seed increased with increasing soil depth. Fleabane was also found to be sensitive to soil type with no seedlings emerging in the self-mulching black vertisol soil. A strategic tillage trial showed that burial of fleabane seed, using a disc or chisel plough, to a depth of greater than 2 cm can significantly reduce subsequent fleabane emergence. In contrast, tillage increased barnyard grass emergence and tended to decrease persistence. This research showed that weed management plans can not be blanketed across all weed species, rather they need to be targeted for each main weed species.This project has also resulted in an increased knowledge of how to manage fleabane from the eight experiments; one in wheat, two in sorghum, one in cotton and three in fallow on double knock. For summer crops, the best option is to apply a highly effective fallow treatment prior to sowing the crops. For winter crops, the strategy is the integration of competitive crops, residual herbicide followed by a knockdown to control survivors. This project explored further the usefulness of the double knock tactic for weed control and preventing seed set. Two field and one pot experiments have shown that this tactic was highly effective for fleabane control. Paraquat products provided good control when followed by glyphosate. When 2, 4-D was added in a tank mix with glyphosate and followed by paraquat products, 99-100% control was achieved in all cases. The ideal follow-up times for paraquat products after glyphosate were 5-7 days. The preferred follow-up times for 2, 4-D after glyphosate were on the same day and one day later. The pot trial, which compared a population from a cropping field with previous glyphosate exposure and a population from a non-cropping area with no previous glyphosate herbicide exposure, showed that the pervious herbicide exposure affected the response of fleabane to herbicidal control measures. The web-based brochure on managing fleabane has been updated.Knowledge on management of summer grasses and safe use of residual herbicides was derived from eight field and pot experiments. Residual grass and broadleaf weed control was excellent with atrazine pre-plant and at-planting treatments, provided rain was received within a short interval after application. Highly effective fallow treatments (cultivation and double knock), not only gave excellent grass control in the fallow, also gave very good control in the following cotton. In the five re-cropping experiments, there were no adverse impacts on cotton from atrazine, metolachlor, metsulfuron and chlorsulfuron residues following use in previous sorghum, wheat and fallows. However, imazapic residues did reduce cotton growth.The development of strategies to reduce the heavy reliance on glyphosate in our cropping systems, and therefore minimise the risk of glyphosate resistance development, was a key factor in the research undertaken. This work included identifying suitable tactics for summer grass control, such as double knock with glyphosate followed by paraquat and tillage. Research on fleabane also concentrated on minimising emergence through tillage, and applying the double knock tactic. Our studies have shown that these strategies can be used to prevent seed set with the goal of driving down the seed bank. Utilisation of the strategies will also reduce the reliance on glyphosate, and therefore reduce the risk of glyphosate resistance developing in our cropping systems.Information from this research, including ecological and management data were collected from an additional eight paddock monitoring sites, was also incorporated into the Weeds CRC seed bank model "Weed Seed Wizard", which will be able to predict the impact of different management options on weed populations in cotton and grain farming systems. Extensive communication activities were undertaken throughout this project to ensure adoption of the new strategies for improved weed management and reduced risk for glyphosate resistance.

Soil seed bank longevity information for weed eradication target species

The longevity of seed in the soil is a key determinant of the cost and length of weed eradication programs. Soil seed bank information and ongoing research have input into the planning and reporting of two nationally cost shared weed eradication programs based in tropical north Queensland. These eradication programs are targeting serious weeds such as Chromoleana odorata, Mikania micrantha, Miconia calvescens, Clidemia hirta and Limnocharis flava. Various methods are available for estimating soil seed persistence. Field methods to estimate the total and germinable soil seed densities include seed packet burial trials, extracting seed from field soil samples, germinating seed in field soil samples and observations from native range seed bank studies. Interrogating field control records can also indicate the length of the control and monitoring periods needed to exhaust the seed bank. Recently, laboratory tests which rapidly age seed have provided an additional indicator of relative seed persistence. Each method has its advantages, drawbacks and logistical constraints.