Assessment of a Novel Real-Time Bio-Liquor Circulation System for Manure Management and Mitigation of Odor Potential in Swine Farming.

Recently, circulating biologically treated manure in slurry pits has been used as an odor reduction technology, but few successful results have been reported, due to the lack of proper control strategies for bioreactors. This study was conducted to investigate the performance of the developed real-time controlled bio-liquor circulation system (BCS) at farm scale. The BCS was operated sequentially as per swine manure inflow (anoxic, aerobic, and settling) circulation to the slurry pit. Each operational phase was self-adjusted in real-time using a novel algorithm for detecting the control point on the oxidation reduction potential (ORP) and pH (mV)-time profiles, the nitrogen break point (NBP), and the nitrate knee point (NKP) in the aerobic and anoxic phases, respectively. The NH4-N in the slurry manure was thoroughly removed (100%) in the bioreactor, optimizing the duration of each operational phase by accurately detecting real-time control points. The newly developed real-time BCS decreased the nitrogen and organic matter in the slurry pit by >70%, and the potential ammonia and methane emissions by 75% and 95%, respectively. This study highlights that improved BCS that utilizes ORP tracking and pH (mV)-time profiles can effectively optimize BCS operation, and thereby reduce malodor and GHG emissions from swine farms.

Real-Time Control Technology for a Bio-Liquor Circulation System in a Swine Barn with Slurry Pit: Pilot Scale Study.

The livestock industry, especially swine production, has been pressurized by vicinity complaints about odor in Korea. Therefore, a lot of effort has been undertaken regarding reducing the odor emissions from pigsties, widely carried out and the washing out manure in slurry pit by liquid-phase compost has particularly been spotlighted with outstanding performance of odor reduction. However, such a washing out manure called bio-liquor circulation system (BCS) has been controlled by a timer with designated reaction time, which cannot guarantee the system performance. This research proposes an effective real-time control technology for BCS, which circulates bio-liquor to the slurry pit of swine barns. The real-time control system was operated through accurate detection of the designated control points on the oxidation reduction potential (ORP) and pH time profiles for the nitrate knee point (NKP) and nitrogen break point (NBP) in anoxic and aerobic conditions with 100 and 99.6% performances, respectively. The duration of the anoxic and aerobic phases was also automated and noticeably lowered the concentration of nutrients in the manure in the slurry-pit, which served as a source of malodor. The real-time control strategy may be an innovative way to reduce odor and simultaneously produce liquid fertilizer, and provides a reference for the optimization of the industrial scale.

Nutrient recovery from swine wastewater at full-scale: An integrated technical, economic and environmental feasibility assessment.

In this study, the technical, economic and environmental attributes of a full-scale nutrient recovery process connected to the centralized swine wastewater treatment facility (CSWTF) were evaluated. The performance of the process was assessed by introducing influent to the recovery reactor from different components of the CSWTF such as sedimentation tank (swine wastewater) and biological treatment reactor (biologically oxidized material and supernatant of the biologically oxidized material). The results of technical performance assessment revealed that the O-P recovery (87.1-90.7%) and NH4-N removal (66.9-72.1%) efficiencies from the influent of biological treatment reactor were significantly higher than the influent from sedimentation tank (81.7 and 19.8%, respectively, p < 0.05). The economic evaluation elucidated that by increasing the treatment capacity of the recovery reactor from 30 m3/d to 100 m3/d, operating expenses could be covered through the commercialization of struvite, while it would take around seven years to get back the capital investment. Additional economic savings could also be possible when using the recovered struvite as a fertilizer raw material along with other environmental benefits. Considering the current farming practices in Korea, the complete recovery of O-P from CSWTFs as struvite could drop the soil phosphorus surplus by 40%, minimize the phosphatic fertilizer consumption by 6.4% and ultimately reduce CO2 equivalent emissions of 6522 tons/year in comparison to chemical fertilizer production. However, during the continuous operation of the full-scale nutrient recovery process, influent characteristics need to be incessantly monitored and adjusted to the optimum conditions to improve the economics of recovered products. Overall, the nutrient recovery process at full-scale not only solves the problem of treating highly polluted swine wastewater but also helps to ensure societal and environmental sustainability.

Development of a Real-Time Controlled Bio-Liquor Circulation System for Swine Farms: A Lab-Scale Study.

In this study, an attempt was made to develop a real-time control strategy using oxidation-reduction potential (ORP) and pH (mV) time profiles for the efficient operation of bio-liquor circulation system (BCS) in swine farms and its effectiveness in reducing odor emission through improving manure properties in the slurry pit was evaluated. The lab-scale BCS used in this study comprised a bioreactor and a slurry pit. The bioreactor was operated in a sequence of inflow of swine manure ® anoxic phase ® aerobic phase ® circulation to the slurry pit. The improvement in swine manure properties was elucidated by comparing the results of the BCS slurry pit (circulation type, CT) and conventional slurry pit (non-circulation type, NCT). The results revealed that the ORP time profile successfully detected the nitrate knee point (NKP) in the anoxic phase. However, it was less stable in detecting the nitrogen break point (NBP) in the aerobic phase. The pH (mV) time profile showed a more efficient detection of NBP. Compared to the NCT slurry pit, concentrations of ammonium nitrogen (NH4-N) and soluble total organic carbon (STOC) and other analyzed swine manure properties were much lower in the CT slurry pit. In the aspect of odor reduction, around 98.3% of NH3 was removed in the CT slurry pit. The real-time controlled BCS can overcome the drawbacks of fixed time-based BCS operation and therefore can be considered as a useful tool to reduce odor emission from intensive swine farming operations. However, further studies and refinement in control algorithms might be required prior to its large-scale application.

Simultaneous Removal of Pollutants and Recovery of Nutrients from High-Strength Swine Wastewater Using a Novel Integrated Treatment Process.

In this study, a novel treatment approach combining biological treatment, struvite crystallization, and electrochemical treatment was developed and its efficiency for the simultaneous removal of pollutants and recovery of nutrients from high strength swine wastewater (SWW) was verified. For all the parameters, maximum removal efficiencies in the lab-scale test were obtained in the range of 93.0-98.7% except for total solids (TS) (79.4%). Farm-scale process showed overall removal efficiencies for total nitrogen (TN), total phosphorus (TP), soluble total organic carbon (sTOC), and color as 94.5%, 67.0%, 96.1%, and 98.9%, respectively, while TS, suspended solids (SS), ammonium nitrogen (NH4-N), and ortho-phosphate (O-P) concentrations were reduced by 91.5%, 99.6%, 98.6%, and 91.9%, respectively. Moreover, the struvite recovered from SWW showed heavy metal concentrations within the range of the Korean standard for fertilizers and feedstocks and thus, suggesting its potential application as fertilizer and in animal feed production. Using the proposed process, the SWW was converted to liquid compost as a quick-acting fertilizer, struvite as a slow-release fertilizer, and the decolorized and disinfected effluent after electrochemical treatment was safe for discharge according to Korean standard. Therefore, the novel integrated treatment process used in this study can be considered as a solution for SWW management and for the simultaneous removal and recycling of nutrients (N and P).

Rational budgeting approach as a nutrient management tool for mixed crop-swine farms in Korea.

OBJECTIVE: Due to rapid economic return, mixed crop-swine farming systems in Korea have become more intensive. Intensive farming practices often cause nutrient surpluses and lead to environmental pollution. Nutrient budgets can be used to evaluate the environmental impact and as a regulatory policy instrument for nutrient management. This study was conducted to select a nutrient budgeting approach applicable to the mixed crop-swine farms in Korea and suggest an effective manure treatment method to reduce on-farm nutrient production. METHODS: In this study, we compared current and ideal gross nutrient balance (GNB) approaches of Organisation for Economic Co-operation and Development and soil system budget (SSB) approach with reference to on-farm manure treatment processes. Data obtained from farm census and published literature were used to develop the farm nutrient budgets. RESULTS: The average nitrogen (N) and phosphorus (P) surpluses were approximately 11 times and over 7 times respectively higher in the GNB approaches than the SSB. After solid-liquid separation of manure, during liquid composting a change in aeration method from intermittent to continuous reduced the N and P loading about 50% and 47%, respectively. Although changing in solid composting method from turning only to turning+aeration improved the N removal efficiency by 30.5%, not much improvement in P removal efficiency was observed. CONCLUSION: Although the GNB approaches depict the impact of nutrients produced in the mixed crop-swine farms on the overall agricultural environment, the SSB approach shows the partitioning among different nutrient loss pathways and storage of nutrients within the soil system; thus, can help design sustainable nutrient management plans for the mixed cropswine farms. The study also suggests that continuous aeration for liquid composting and turning+aeration for solid composting can reduce nutrient loading to the soil.

In Vivo Toxicity and In Vitro Solubility Assessment of Pre-Treated Struvite as A Potential Alternative Phosphorus Source in Animal Feed.

Apart from using as fertilizer for plants, the application of struvite may be expanded to animal feed industries through proper pre-treatment. This study aimed to investigate the safety and efficacy of using pre-treated struvite (microwave irradiated struvite (MS) and incinerated struvite (IS)) in animal feeds. For safety assessment, an in vivo toxicity experiment using thirty female Sprague Dawley rats (average body weight (BW) of 200 ± 10 g) was conducted. The rats were randomly divided into five groups, including a control. Based on the BW, MS and IS were applied daily by oral administration with 1 and 10 mg kg-1-BW (MS1 and MS10; IS1 and IS10) using dimethyl sulfoxide (DMSO) as a vehicle. A series of jar tests were conducted for four hours to check the solubility of the MS and IS at different pH (pH 2, 4, and 5) and compared to a commercial P source (monocalcium phosphate, MCP, control). The toxicity experiment results showed no significant differences among the treatments in BW and organ (liver, kidney, heart, and lung) weight of rats (p > 0.05). There were no adverse effects on blood parameters and the histopathological examination showed no inflammation in the organ tissues in MS and IS treated groups compared to the control. In an in vitro solubility test, no significant difference was observed in ortho-phosphate (O-P) solubility from the MCP and MS at pH 2 and 4 (p > 0.05), while O-P solubility from MS at pH 5 to 7 was higher than MCP and found to be significantly different (p < 0.05). O-P solubility from IS was the lowest among the treatments and significantly different from MCP and MS in all the experiments (p < 0.05). The results of this study not only suggest that the struvite pre-treated as MS could be a potential alternative source of P in animal feed but also motivate further studies with more stringent designs to better examine the potential of struvite application in diverse fields.

Nutrient production from Korean poultry and loading estimations for cropland.

BACKGROUND: Poultry breeding has increased by 306% in Korea, inevitably increasing the production of manure which may contribute to environmental pollution. The nutrients (NP) in the manure are essential for crop cultivation and soil fertility when applied as compost. Excess nutrients from manure can be accumulated on the land and can lead to eutrophication. Therefore, a nutrient load on the finite land should be calculated. METHODS: This study calculates the nutrient production from Korean poultry by investigating 11 broiler and 16 laying hen farms. The broiler manure was composted using deep litter composting while for layer deep litter composting, drying, and simple static pile were in practice. The effect of weight reduction and storing period during composting was checked. Three weight reduction cases of compost were constructed to calculate nutrient loading coefficients (NLCs) using data from; i) farm investigation, ii) theoretical P changes (ΔP = 0), and iii) dry basis. RESULTS: During farm investigation of broiler and layer with deep litter composting, there was a 68 and 21% N loss whereas 77 and 33% P loss was found, respectively. In case of layer composting, a loss of 10-56% N and a 52% P loss was observed. Drying manure increased the P concentrations therefore NLCs calculated using dry basis that showed quite higher reductions (67% N; 53% P). Nutrient loss from farm investigation was much higher than reported by Korean Ministry of Environment (ME). CONCLUSIONS: Nutrients in manure are decreased when undergo storing or composting process due to microbial action, drying, and leaching. The nutrient load applied to soil is less than the fresh manure, hence the livestock manure management and conservation of environment would be facilitated.

Nutrient variations from swine manure to agricultural land.

OBJECTIVE: Swine manure in Korea is separated into solid and liquid phases which are composted separately and then applied on land. The nutrient accumulation in soil has been a big issue in Korea but the basic investigation about nutrient input on arable land has not been achieved in detail. Within the nutrient production from livestock at the national level, most values are calculated by multiplication of the number of animals with the excreta unit per animal. However, the actual amount of nutrients from swine manure may be totally different with the nutrients applied to soil since livestock breeding systems are not the same with each country. METHODS: This study investigated 15 farms producing solid compost and 14 farms producing liquid compost. Composting for solid phase used the Turning+Aeration (TA) or Turning (T) only methods, while liquid phase aeration composting was achieved by continuous (CA), intermittent (IA), or no aeration (NA). Three scenarios were constructed for investigating solid compost: i) farm investigation, ii) reference study, and iii) theoretical P changes (ΔP = 0), whereas an experiment for water evaporation was conducted for analyzing liquid compost. RESULTS: In farm investigation, weight loss rates of 62% and 63% were obtained for TA and T, respectively, while evaporation rates for liquid compost were 8.75, 7.27, and 5.14 L/m2·d for CA, IA, and NA, respectively. Farm investigation provided with the combined nutrient load (solid+liquid) of VS, N, and P of 117.6, 7.2, and 2.7 kg/head·yr. Nutrient load calculated from farm investigation is about two times higher than the calculated with reference documents. CONCLUSION: The nutrient loading coefficients from one swine (solid+liquid) were (volatile solids, 0.79; nitrogen, 0.53; phosphorus, 0.71) with nutrient loss of 21%, 47%, and 29%, respectively. The nutrient count from livestock manure using the excretion unit has probably been overestimated without consideration of the nutrient loss.

Nutrient production from dairy cattle manure and loading on arable land.

OBJECTIVE: Along with increasing livestock products via intensive rearing, the accumulation of livestock manure has become a serious issue due to the fact that there is finite land for livestock manure recycling via composting. The nutrients from livestock manure accumulate on agricultural land and the excess disembogues into streams causing eutrophication. In order to systematically manage nutrient loading on agricultural land, quantifying the amount of nutrients according to their respective sources is very important. However, there is a lack of research concerning nutrient loss from livestock manure during composting or storage on farms. Therefore, in the present study we quantified the nutrients from dairy cattle manure that were imparted onto agricultural land. METHODS: Through investigation of 41 dairy farms, weight reduction and volatile solids (VS), total nitrogen (TN), and total phosphorus (TP) changes of dairy cattle manure during the storage and composting periods were analyzed. In order to support the direct investigation and survey on site, the three cases of weight reduction during the storing and composting periods were developed according to i) experiment, ii) reference, and iii) theoretical changes in phosphorus content (ΔP = 0). RESULTS: The data revealed the nutrient loading coefficients (NLCs) of VS, TN, and TP on agricultural land were 1.48, 0.60, and 0.66, respectively. These values indicated that the loss of nitrogen and phosphorus was 40% and 34%, respectively, and that there was an increase of VS since bedding materials were mixed with excretion in the barn. CONCLUSION: As result of nutrient-footprint analyses, the amounts of TN and TP particularly entered on arable land have been overestimated if applying the nutrient amount in fresh manure. The NLCs obtained in this study may assist in the development of a database to assess the accurate level of manure nutrient loading on soil and facilitate systematic nutrient management.

Structure of cellular ESCRT-III spirals and their relationship to HIV budding.

The ESCRT machinery along with the AAA+ ATPase Vps4 drive membrane scission for trafficking into multivesicular bodies in the endocytic pathway and for the topologically related processes of viral budding and cytokinesis, but how they accomplish this remains unclear. Using deep-etch electron microscopy, we find that endogenous ESCRT-III filaments stabilized by depleting cells of Vps4 create uniform membrane-deforming conical spirals which are assemblies of specific ESCRT-III heteropolymers. To explore functional roles for ESCRT-III filaments, we examine HIV-1 Gag-mediated budding of virus-like particles and find that depleting Vps4 traps ESCRT-III filaments around nascent Gag assemblies. Interpolating between the observed structures suggests a new role for Vps4 in separating ESCRT-III from Gag or other cargo to allow centripetal growth of a neck constricting ESCRT-III spiral.

COPII and the regulation of protein sorting in mammals.

Secretory proteins are transported to the Golgi complex in vesicles that bud from the endoplasmic reticulum. The cytoplasmic coat protein complex II (COPII) is responsible for cargo sorting and vesicle morphogenesis. COPII was first described in Saccharomyces cerevisiae, but its basic function is conserved throughout all eukaryotes. Nevertheless, the COPII coat has adapted to the higher complexity of mammalian physiology, achieving more sophisticated levels of secretory regulation. In this review we cover aspects of mammalian COPII-mediated regulation of secretion, in particular related to the function of COPII paralogues, the spatial organization of cargo export and the role of accessory proteins.

Cell biology of the ESCRT machinery.

The ESCRT (endosomal sorting complex required for transport) machinery comprises a set of protein complexes that regulate sorting and trafficking into multivesicular bodies en route to the lysosome. The physical mechanism responsible for generating lumenal vesicles in this pathway is unknown. Here we review recent studies suggesting that components of the ESCRT-III complex drive lumenal vesicle formation and consider possible mechanisms for this reaction.

Novel interactions of ESCRT-III with LIP5 and VPS4 and their implications for ESCRT-III disassembly.

The AAA+ ATPase VPS4 plays an essential role in multivesicular body biogenesis and is thought to act by disassembling ESCRT-III complexes. VPS4 oligomerization and ATPase activity are promoted by binding to LIP5. LIP5 also binds to the ESCRT-III like protein CHMP5/hVps60, but how this affects its function remains unclear. Here we confirm that LIP5 binds tightly to CHMP5, but also find that it binds well to additional ESCRT-III proteins including CHMP1B, CHMP2A/hVps2-1, and CHMP3/hVps24 but not CHMP4A/hSnf7-1 or CHMP6/hVps20. LIP5 binds to a different region within CHMP5 than within the other ESCRT-III proteins. In CHMP1B and CHMP2A, its binding site encompasses sequences at the proteins' extreme C-termini that overlap with "MIT interacting motifs" (MIMs) known to bind to VPS4. We find unexpected evidence of a second conserved binding site for VPS4 in CHMP2A and CHMP1B, suggesting that LIP5 and VPS4 may bind simultaneously to these proteins despite the overlap in their primary binding sites. Finally, LIP5 binds preferentially to soluble CHMP5 but instead to polymerized CHMP2A, suggesting that the newly defined interactions between LIP5 and ESCRT-III proteins may be regulated by ESCRT-III conformation. These studies point to a role for direct binding between LIP5 and ESCRT-III proteins that is likely to complement LIP5's previously described ability to regulate VPS4 activity.

CHMP4B, a novel gene for autosomal dominant cataracts linked to chromosome 20q.

Cataracts are a clinically diverse and genetically heterogeneous disorder of the crystalline lens and a leading cause of visual impairment. Here we report linkage of autosomal dominant "progressive childhood posterior subcapsular" cataracts segregating in a white family to short tandem repeat (STR) markers D20S847 (LOD score [Z] 5.50 at recombination fraction [theta] 0.0) and D20S195 (Z=3.65 at theta =0.0) on 20q, and identify a refined disease interval (rs2057262-(3.8 Mb)-rs1291139) by use of single-nucleotide polymorphism (SNP) markers. Mutation profiling of positional-candidate genes detected a heterozygous transversion (c.386A-->T) in exon 3 of the gene for chromatin modifying protein-4B (CHMP4B) that was predicted to result in the nonconservative substitution of a valine residue for a phylogenetically conserved aspartic acid residue at codon 129 (p.D129V). In addition, we have detected a heterozygous transition (c.481G-->A) in exon 3 of CHMP4B cosegregating with autosomal dominant posterior polar cataracts in a Japanese family that was predicted to result in the missense substitution of lysine for a conserved glutamic acid residue at codon 161 (p.E161K). Transfection studies of cultured cells revealed that a truncated form of recombinant D129V-CHMP4B had a different subcellular distribution than wild type and an increased capacity to inhibit release of virus-like particles from the cell surface, consistent with deleterious gain-of-function effects. These data provide the first evidence that CHMP4B, which encodes a key component of the endosome sorting complex required for the transport-III (ESCRT-III) system of mammalian cells, plays a vital role in the maintenance of lens transparency.

Structure/function analysis of four core ESCRT-III proteins reveals common regulatory role for extreme C-terminal domain.

Endosomal sorting complex required for transport-III (ESCRT-III) is a large complex built from related ESCRT-III proteins involved in multivesicular body biogenesis. Little is known about the structure and function of this complex. Here, we compare four human ESCRT-III proteins - hVps2-1/CHMP2a, hVps24/CHMP3, hVps20/CHMP6, and hSnf7-1/CHMP4a - to each other, studying the effects of deleting predicted alpha-helical domains on their behavior in transfected cells. Surprisingly, removing approximately 40 amino acids from the C-terminus of each protein unmasks a common ability to associate with endosomal membranes and assemble into large polymeric complexes. Expressing these truncated ESCRT-III proteins in cultured cells causes ubiquitinated cargo to accumulate on enlarged endosomes and inhibits viral budding, while expressing full-length proteins does not. hVps2-1/CHMP2a lacking its C-terminal 42 amino acids further fails to bind to the AAA+ adenosine triphosphatase VPS4B/SKD1, indicating that C-terminal sequences are important for interaction of ESCRT-III proteins with VPS4. Overall, our study supports a model in which ESCRT-III proteins cycle between a default 'closed' state and an activated 'open' state under control of sequences at their C-terminus and associated factors.

Mouse model of N-acetylgalactosamine-6-sulfate sulfatase deficiency (Galns-/-) produced by targeted disruption of the gene defective in Morquio A disease.

Mucopolysaccharidosis IVA is an autosomal recessive disorder caused by a deficiency of N-acetylgalactosamine-6-sulfate sulfatase (GALNS), a lysosomal enzyme required for the stepwise degradation of keratan sulfate (KS) and chondroitin-6-sulfate (C6S). To generate a model for studies of the pathophysiology and of potential therapies, we disrupted exon 2 of Galns, the homologous murine gene. Homozygous Galns-/- mice have no detectable GALNS enzyme activity and show increased urinary glycosaminoglycan (GAGs) levels. These mice accumulate GAGs in multiple tissues including liver, kidney, spleen, heart, brain and bone marrow. At 2 months old, lysosomal storage is present primarily within reticuloendothelial cells such as Kupffer cells and cells of the sinusoidal lining of the spleen. Additionally, by 12 months old, vacuolar change is observed in the visceral epithelial cells of glomeruli and cells at the base of heart valves but it is not present in parenchymal cells such as hepatocytes and renal tubular epithelial cells. In the brain, hippocampal and neocortical neurons and meningeal cells had lysosomal storage. KS and C6S were more abundant in the cytoplasm of corneal epithelial cells of Galns-/- mice compared with wild-type mice by immunohistochemistry. Radiographs revealed no change in the skeletal bones of mice up to 12 months old. Thus, targeted disruption of the murine Galns gene has produced a murine model, which shows visceral storage of GAGs but lacks the skeletal features. The complete absence of GALNS in mutant mice makes them useful for studies of pharmacokinetics and tissue targeting of recombinant GALNS designed for enzyme replacement.