Fate of pathogen indicators in a domestic blend of food waste and wastewater through a two-stage anaerobic digestion system

Anaerobic digestion is a viable on-site treatment technology for rich organic waste streams such as food waste and blackwater. In contrast to large-scale municipal wastewater treatment plants which are typically located away from the community, the effluent from any type of on-site system is a potential pathogenic hazard because of the intimacy of the system to the community. The native concentrations of the pathogen indicators Escherichia coli, Clostridium perfringens and somatic coliphage were tracked for 30 days under stable operation (organic loading rate (OLR) = 1.8 kgCOD m(-3) day(-1), methane yield = 52% on a chemical oxygen demand (COD) basis) of a two-stage laboratory-scale digester treating a mixture of food waste and blackwater. E. coli numbers were reduced by a factor of 10(6.4) in the thermophilic stage, from 10(7.5+/-0.3) to 10(1.1+/-0.1) cfu 100 mL(-1), but regenerated by a factor of 10(4) in the mesophilic stage. Neither the thermophilic nor mesophilic stages had any significant impact on C. perfringens concentrations. Coliphage concentrations were reduced by a factor of 10(1.4) across the two stages. The study shows that anaerobic digestion only reduces pathogen counts marginally but that counts in effluent samples could be readily reduced to below detection limits by filtration through a 0.22 microm membrane, to investigate membrane filtration as a possible sanitation technique.

Validation of PigBal model predictions for pig manure production

PigBal is a mass balance model that uses pig diet, digestibility and production data to predict the manure solids and nutrients produced by pig herds. It has been widely used for designing piggery effluent treatment systems and sustainable reuse areas at Australian piggeries. More recently, PigBal has also been used to estimate piggery volatile solids production for assessing greenhouse gas emissions for statutory reporting purposes by government, and for evaluating the energy potential from anaerobic digestion of pig effluent. This paper has compared PigBal predictions of manure total, volatile, and fixed solids, and nitrogen (N), phosphorus (P) and potassium (K), with manure production data generated in a replicated trial, which involved collecting manure from pigs housed in metabolic pens. Predictions of total, volatile, and fixed solids and K in the excreted manure were relatively good (combined diet R2 ≥ 0.79, modelling efficiency (EF) ≥ 0.70) whereas predictions of N and P, were generally less accurate (combined diet R2 0.56 and 0.66, EF 0.19 and –0.22, respectively). PigBal generally under-predicted lower N values while over-predicting higher values, and generally over-predicted manure P production for all diets. The most likely causes for this less accurate performance were ammonium-N volatilisation losses between manure excretion and sample analysis, and the inability of PigBal to account for higher rates of P uptake by pigs fed diets containing phytase. The outcomes of this research suggest that there is a need for further investigation and model development to enhance PigBal’s capabilities for more accurately assessing nutrient loads. However, PigBal’s satisfactory performance in predicting solids excretion demonstrates that it is suitable for assessing the methane component of greenhouse gas emission and the energy potential from anaerobic digestion of volatile solids in piggery effluent. The apparent overestimation of N and P excretion may result in conservative nutrient application rates to land and the over-prediction of the nitrous oxide component of greenhouse gas emissions.

Validation of PigBal model predictions for pig manure production

PigBal is a mass balance model that uses pig diet, digestibility and production data to predict the manure solids and nutrients produced by pig herds. It has been widely used for designing piggery effluent treatment systems and sustainable reuse areas at Australian piggeries. More recently, PigBal has also been used to estimate piggery volatile solids production for assessing greenhouse gas emissions for statutory reporting purposes by government, and for evaluating the energy potential from anaerobic digestion of pig effluent. This paper has compared PigBal predictions of manure total, volatile, and fixed solids, and nitrogen (N), phosphorus (P) and potassium (K), with manure production data generated in a replicated trial, which involved collecting manure from pigs housed in metabolic pens. Predictions of total, volatile, and fixed solids and K in the excreted manure were relatively good (combined diet R2 ≥ 0.79, modelling efficiency (EF) ≥ 0.70) whereas predictions of N and P, were generally less accurate (combined diet R2 0.56 and 0.66, EF 0.19 and -0.22, respectively). PigBal generally under-predicted lower N values while over-predicting higher values, and generally over-predicted manure P production for all diets. The most likely causes for this less accurate performance were ammonium-N volatilisation losses between manure excretion and sample analysis, and the inability of PigBal to account for higher rates of P uptake by pigs fed diets containing phytase. The outcomes of this research suggest that there is a need for further investigation and model development to enhance PigBal's capabilities for more accurately assessing nutrient loads. However, PigBal's satisfactory performance in predicting solids excretion demonstrates that it is suitable for assessing the methane component of greenhouse gas emission and the energy potential from anaerobic digestion of volatile solids in piggery effluent. The apparent overestimation of N and P excretion may result in conservative nutrient application rates to land and the over-prediction of the nitrous oxide component of greenhouse gas emissions. © CSIRO 2016.

Odour emissions from anaerobic piggery ponds. 2: Improving estimates of emission rate through recognition of spatial variability

Odour emission rates were measured for seven different anaerobic ponds treating piggery wastes at six to nine discrete locations across the surface of each pond on each sampling occasion over a thirteen month period. Significant variability in emission rates were observed for each pond. Measurement of a number of water quality variables in pond liquor samples collected at the same time and from the same locations as the odour samples indicated that the composition of the pond liquor was also variable. The results indicated that spatial variability was a real phenomenon and could have a significant impact on odour assessment practices. Considerably more odour samples would be required to characterise pond emissions than currently recommended by most practitioners, or regulatory agencies.

Odour emissions from anaerobic piggery ponds. 1. Results of a three season, 14-month survey

Odour emission rates were measured for seven different anaerobic ponds treating piggery wastes at six to nine discrete locations across the surface of each pond on each sampling occasion over a 14-month period. Emission rate values varied between ponds, between seasons for the same pond and even for the same pond on different days of a sampling week. Average seasonal emission rates ranged from 7.9 to 46.5 OU/m2 s, while average emission rates ranged from 16.0 to 29.0 OU/m2 s. Factors potentially responsible for the variability in emission rates were investigated, including air and pond liquor temperatures, time of day of sample collection, season and the impact of a prolonged drought.

Lack of release of bound anthocyanins and phenolic acids from carrot plant cell walls and model composites during simulated gastric and small intestinal digestion

Separately, polyphenols and plant cell walls (PCW) are important contributors to the health benefits associated with fruits and vegetables. However, interactions with PCW which occur either during food preparation or mastication may affect bioaccessibility and hence bioavailability of polyphenols. Binding interactions between anthocyanins, phenolic acids (PAs) and PCW components, were evaluated using both a bacterial cellulose-pectin model system and a black carrot puree system. The majority of available polyphenols bound to PCW material with 60-70% of available anthocyanins and PAs respectively binding to black carrot puree PCW matter. Once bound, release of polyphenols using acidified methanol is low with only similar to 20% of total anthocyanins to similar to 30% of PAs being released. Less than 2% of bound polyphenol was released after in vitro gastric and small intestinal (S.I.) digestion for both the model system and the black carrot puree PCW matter. Confocal laser scanning microscopy shows localised binding of anthocyanins to PCW. Very similar patterns of binding for anthocyanins and PAs suggest that PAs form complexes with anthocyanins and polysaccharides. Time dependent changes in extractability with acidified methanol but not the total bound fraction suggests that initial nonspecific deposition on cellulose surfaces is followed by rearrangement of the bound molecules. Minimal release of anthocyanins and PAs after simulated gastric and S.I. digestion indicates that polyphenols in fruits and vegetables which bind to the PCW will be transported to the colon where they would be expected to be released by the action of cell wall degrading bacteria.

QTL associated with barley (Hordeum vulgare) feed quality traits measured through in situ digestion

Barley (Hordeum vulgare) is a major feed source for the intensive livestock industry. Competitiveness against other cereal grains depends largely on the price per unit of expressed feed quality. The traits which contribute to feed quality in barley are largely quantitative in nature but little is known about their genetic control. A study to identify the quantitative trait loci (QTLs) associated with feed quality was performed using a F6-derived recombinant inbred barley population. Samples from each line were incubated in the rumen of fistulated cattle, recovered, washed and dried for determination of in situ dry matter digestibility. Additionally, both pre- and post-digestion samples were analysed to quantify the content of key quality components relating to acid detergent fibre, total starch and protein. The data was used to identify trait-associated QTLs. Genetic analysis identified significant QTLs on chromosomes 2H, 5H and 7H. Genetic markers linked to these QTL should provide an effective tool for the selection and improvement of feed barley in the future.

Digestion of forages in the rumen is increased by the amount but not the type of protein supplement

Three polyester bag experiments were conducted with fistulated Bos indicus steers to determine the effect of the amount and type of nitrogen (N) supplement on the digestion rate of forages different in quality. In Experiment 1, test substrates were incubated in polyester bags in the rumen of steers fed ryegrass, pangola grass, speargrass and Mitchell grass hays in a 4 by 4 Latin-square design. In Experiment 2, test substrates were incubated in polyester bags in the rumen of steers fed speargrass hay supplemented with urea and ammonium sulfate (US), branched-chain amino acids with US (USAA), casein, cottonseed meal, yeast and Chlorella algae in a 7 by 3 incomplete Latin-square design. In Experiment 3, test substrates were incubated in polyester bags in the rumen of steers fed Mitchell grass hay supplemented with increasing amounts of US or Spirulina algae (Spirulina platensis). The test substrates used in all experiments were speargrass, Mitchell grass, pangola grass or ryegrass hays. Digestion rate of the ryegrass substrate was higher than that of the speargrass substrate (P < 0.05) in Experiment 1. Supplementation with various N sources increased the degradation rate and effective degradability of all incubated substrates above that apparent in Control steers (P < 0.05; Experiment 2). Supplementation of US and Spirulina increased degradation rate and effective degradability of ryegrass, pangola grass and Mitchell grass substrates above that apparent in Control steers (P < 0.05; Experiment 3). However, there was no further response on digestion rate of the substrates in increasing supplementation levels either for US or Spirulina. In conclusion, rate of digestion was affected by forage physical and anatomical properties. Supplementation with various N sources increased rate of digestion when the Control forage ration was very low in N but once a minimum level of N supplementation was reached, irrespective of form of N or other potential growth factors, there was no further increase in rate of digestion.

Effect of varying the proportion of molasses in the diet on intake, digestion and microbial protein production by steers

The present experiment was conducted to determine the efficiency of microbial protein production in the rumen and intake by cattle fed high-molasses diets. Intake and microbial crude protein (MCP) production were measured along with the concentration of rumen ammonia-nitrogen (N) and volatile fatty acids (VFA), pH and the rate of digestion of roughage in the rumen. Eight Brahman crossbred steers weighing 211 ± 19.3 (± s.d.) kg were used in a double 4 × 4 Latin square design. Steers were allocated to one of four total mixed rations: control (pangola hay only), 25M (25% molasses/urea mix + 75% hay), 50M (50% molasses/urea + 50% hay), and 75M (75% molasses/urea + 25% hay). The production and efficiency of production of MCP (EMCP) of the diet increased quadratically as the level of molasses in the diet increased. The EMCP from the molasses/urea mix was estimated as 166 g MCP/kg digestible organic matter (DOM), a relatively high value. Intake of dry matter (DM) and DOM increased quadratically, reaching a peak when molasses was ~50% (as fed) of the ration. Digestibility of DM increased quadratically and that of neutral detergent fibre decreased linearly with increasing level of molasses in the diet. Molasses inclusion in the diet had no effect on rumen pH, ammonia and VFA concentration in the rumen fluid, plasma urea-N, urine pH or ruminal fractional outflow rate of ytterbium-labelled particles and Cr-EDTA. It was concluded that a diet with a high level of molasses (>50%) and supplemented with adequate N had high EMCP, and that low MCP production was not a factor limiting intake or performance of cattle consuming high-molasses diets.

Effect of varying the proportion of molasses in the diet on intake, digestion and microbial protein production by steers

The present experiment was conducted to determine the efficiency of microbial protein production in the rumen and intake by cattle fed high-molasses diets. Intake and microbial crude protein (MCP) production were measured along with the concentration of rumen ammonia-nitrogen (N) and volatile fatty acids (VFA), pH and the rate of digestion of roughage in the rumen. Eight Brahman crossbred steers weighing 211 ± 19.3 (± s.d.) kg were used in a double 4 x 4 Latin square design. Steers were allocated to one of four total mixed rations: control (pangola hay only), 25M (25% molasses/urea mix + 75% hay), 50M (50% molasses/urea + 50% hay), and 75M (75% molasses/urea + 25% hay). The production and efficiency of production of MCP (EMCP) of the diet increased quadratically as the level of molasses in the diet increased. The EMCP from the molasses/urea mix was estimated as 166 g MCP/kg digestible organic matter (DOM), a relatively high value. Intake of dry matter (DM) and DOM increased quadratically, reaching a peak when molasses was ∼50% (as fed) of the ration. Digestibility of DM increased quadratically and that of neutral detergent fibre decreased linearly with increasing level of molasses in the diet. Molasses inclusion in the diet had no effect on rumen pH, ammonia and VFA concentration in the rumen fluid, plasma urea-N, urine pH or ruminal fractional outflow rate of ytterbium-labelled particles and Cr-EDTA. It was concluded that a diet with a high level of molasses (>50%) and supplemented with adequate N had high EMCP, and that low MCP production was not a factor limiting intake or performance of cattle consuming high-molasses diets.

An overview of land application of pig effluent-P using soil P chemistry and mass balance calculations.

Attention is directed at land application of piggery effluent (containing urine, faeces, water, and wasted feed) as a potential source of water resource contamination with phosphorus (P). This paper summarises P-related properties of soil from 0-0.05 m depth at 11 piggery effluent application sites, in order to explore the impact that effluent application has had on the potential for run-off transport of P. The sites investigated were situated on Alfisol, Mollisol, Vertisol, and Spodosol soils in areas that received effluent for 1.5-30 years (estimated effluent-P applications of 100-310000 kg P/ha in total). Total (PT), bicarbonate extractable (PB), and soluble P forms were determined for the soil (0-0.05 m) at paired effluent and no-effluent sites, as well as texture, oxalate-extractable Fe and Al, organic carbon, and pH. All forms of soil P at 0-0.05 m depth increased with effluent application (PB at effluent sites was 1.7-15 times that at no-effluent sites) at 10 of the 11 sites. Increases in PB were strongly related to net P applications (regression analysis of log values for 7 sites with complete data sets: 82.6 % of variance accounted for, p <0.01). Effluent irrigation tended to increase the proportion of soil PT in dilute CaCl2-extractable forms (PTC: effluent average 2.0 %; no-effluent average 0.6%). The proportion of PTC in non-molybdate reactive forms (centrifuged supernatant) decreased (no-effluent average, 46.4 %; effluent average, 13.7 %). Anaerobic lagoon effluent did not reliably acidify soil, since no consistent relationship was observed for pH with effluent application. Soil organic carbon was increased in most of the effluent areas relative to the no-effluent areas. The four effluent areas where organic carbon was reduced had undergone intensive cultivation and cropping. Current effluent management at many of the piggeries failed to maximise the potential for waste P recapture. Ten of the case-study effluent application areas have received effluent-P in excess of crop uptake. While this may not represent a significant risk of leaching where sorption retains P, it has increased the risk of transport of P by run-off. Where such sites are close to surface water, run-off P loads should be managed.

Piggery pond sludge as a nitrogen source for crops. 1. Mineral N supply estimated from laboratory incubations and field application of stockpiled and wet sludge

Piggery pond sludge (PPS) was applied, as-collected (Wet PPS) and following stockpiling for 12 months (Stockpiled PPS), to a sandy Sodosol and clay Vertosol at sites on the Darling Downs of Queensland. Laboratory measures of N availability were carried out on unamended and PPS-amended soils to investigate their value in estimating supplementary N needs of crops in Australia's northern grains region. Cumulative net N mineralised from the long-term (30 weeks) leached aerobic incubation was described by a first-order single exponential model. The mineralisation rate constant (0.057/week) was not significantly different between Control and PPS treatments or across soil types, when the amounts of initial mineral N applied in PPS treatments were excluded. Potentially mineralisable N (No) was significantly increased by the application of Wet PPS, and increased with increasing rate of application. Application of Wet PPS significantly increased the total amount of inorganic N leached compared with the Control treatments. Mineral N applied in Wet PPS contributed as much to the total mineral N status of the soil as did that which mineralised over time from organic N. Rates of C02 evolution during 30 weeks of aerobic leached incubation indicated that the Stockpiled PPS was more stabilised (19-28% of applied organic C mineralised) than the WetPPS (35-58% of applied organic C mineralised), due to higher lignin content in the former. Net nitrate-N produced following 12 weeks of aerobic non-leached incubation was highly correlated with net nitrate-N leached during 12 weeks of aerobic incubation (R^2 = 0.96), although it was <60% of the latter in both sandy and clayey soils. Anaerobically mineralisable N determined by waterlogged incubation of laboratory PPS-amended soil samples increased with increasing application rate of Wet PPS. Anaerobically minemlisable N from field-moist soil was well correlated with net N mineralised during 30 weeks of aerobic leached incubation (R^2 =0.90 sandy soil; R^2=0.93 clay soil). In the clay soil, the amount of mineral N produced from all the laboratory incubations was significantly correlated with field-measured nitrate-N in the soil profile (0-1.5 m depth) after 9 months of weed-free fallow following PPS application. In contrast, only anaerobic mineralisable N was significantly correlated with field nitrate-N in the sandy soil. Anaerobic incubation would, therefore, be suitable as a rapid practical test to estimate potentially mineralisable N following applications of different PPS materials in the field.

Long-term assessment of efficacy of permeable pond covers for odour reduction

Three anaerobic ponds used to store and treat piggery wastes were fully covered with permeable materials manufactured from polypropylene geofabric, polyethylene shade cloth and supported straw. The covers were assessed in terms of efficacy in reducing odour emission rates over a 40-month period. Odour samples were collected from the surface of the covers, the surface of the exposed liquor and from the surface of an uncovered (control) pond at one of the piggeries. Relative to the emission rate of the exposed liquor at each pond, the polypropylene, shade cloth and straw covers reduced average emission rates by 76%, 69% and 66%, respectively. At the piggery with an uncovered control pond, the polypropylene covers reduced average odour emission rates by 50% and 41%, respectively. A plausible hypothesis, consistent with likely mechanisms for the odour reduction and the olfactometric method used to quantifying the efficacy of the covers, is offered.

Phenotypic variation in natural populations of kikuyu in Australia

The quality of tropical grasses is a major limitation to animal production in tropical and subtropical areas. This is mainly associated with the lower digestibility because C4 grasses have higher fibre levels. Any improvement in quality would require a reduction in the lignin and an increase in the digestion of the neutral detergent fibre content of these plants (Clark and Wilson 1993). Kikuyu (Pennisetum clandestinum) is an important grass for the dairy and beef industries of the subtropics of Australia, South Africa and New Zealand (Mears 1970). Increased digestibility could substantially improve animal production in these industries. These experiments investigated the variation in agronomic and quality of natural populations selected from diverse regions within Australia. Runners of 14 kikuyu selections were collected by project staff or local agronomists from areas considered to have grown kikuyu for over 30 years while Whittet and Noonan were established by seed. Entries were established as single spaced plants on a 1.5 m grid in a randomised block with 3 replicates and evaluated under irrigation at Mutdapilly (brown podsol) and Wollongbar (red ferrosol). Foliage height, forage production and runner yield were assessed along with crude protein (CP), in vitro dry matter digestibility (IVDMD), metabolisable energy (ME), acid detergent fibre (ADF) and neutral detergent fibre (NDF) content of the leaf in autumn, winter and spring.

Assessment of permeable covers for odour reduction in piggery effluent ponds. 1. Laboratory-scale trials

A variety of materials were trialed as supported permeable covers using a series of laboratory-scale anaerobic digesters. Efficacy of cover performance was assessed in terms of impact on odour and greenhouse gas emission rate, and the characteristics of anaerobic liquor. Data were collected over a 12-month period. Initially the covers reduced the rate of odour emission 40-100 times relative to uncovered digesters. After about three months, this decreased to about a threefold reduction in odour emission rate, which was maintained over the remainder of the trial. The covers did not alter methane emission rates. Carbon dioxide emission rates varied according to cover type. Performance of the covers was attributed to the physical characteristics of the cover materials and changes in liquor composition. The reductions in odour emission indicate that these covers offer a cost-effective method for odour control.