Anaerobic composting is a promising method to fully transform food wastes into useful
materials such as biofertilizer and biogas. In this study, the optimum proportions of food
wastes containing vegetable, fruit and meat wastes with dry leaves or cow manure for
composting were determined using the simplex centroid design and response optimizer.
The effectiveness of the pilot-scale composting process was evaluated based on the targeted
compost quality of C/N ratio at 21, pH value at 8 and electrical conductivity of 1 dS/m.
Food wastes composting formulation with dry leaves suggested high percentage of dry
leaves, 86.9% with low food wastes composition of 13.1% constituted by vegetable waste
(1.1%), fruit waste (4.9%) and meat waste (7.1%). With cow manure formulation, only
6% of cow manure was recommended with
another 94.0% of food wastes contributed
by a fair mix of vegetable waste (23.2%),
fruit waste (34.3%) and meat waste (36.5%).
The developed regression models were
experimentally validated with predicted
responses obtained in acceptable ranges for
C/N ratio (21.2 - 21.8), pH (7.92 - 7.99) and
electrical conductivity (0.97 - 1.03 dS/m).
The present study proposes a system for co-composting food waste and poultry manure amended with rice husk biochar at different doses (0, 3, 5, 10%, w/w), saw dust, and salts. The effect of rice husk biochar on the characteristics of final compost was evaluated through stabilization indices such as electrical conductivity, bulk density, total porosity, gaseous emissions and nitrogen conservation. Results indicated that when compared to control, the biochar amendment extended the thermophilic stage of the composting, accelerated the biodegradation and mineralization of substrate mixture and helped in the maturation of the end product. Carbon dioxide, methane and ammonia emissions were reduced and the nitrogen conservation was achieved at a greater level in the 10% (w/w) biochar amended treatments. This study implies that the biochar and salts addition for co-composting food waste and poultry manure is beneficial to enhance the property of the compost.
Antibiotic residues in swine manure when entered the soil would most likely affect the complex composition and functions of the soil microbiome, which is also responsible for degrading these antibiotics. Three different methods of adding ciprofloxacin (CIP), a common antibiotic used in the swine industry, to the soil were used to investigate the effects of CIP on the soil microbiome and the degradation of CIP. Results of the study showed that the microbiome could promote the degradation of CIP in the soil when CIP was incorporated into the soil together with manure. However, the CIP degradation time was prolonged when adding the manure of swine fed with diet containing CIP in the soil. All treatments did not affect the copy number of the resistance genes, except for aac(6')-Ib-cr, as compared with the initial numbers of each treatment. MiSeq Illumina sequencing and Biolog-ECO microplates results showed that CIP had a significant effect on the abundance, structure, and function of the soil microbiome, but different addition methods resulted in distinct effects. Results of the present study demonstrated that the microbiome and fate of CIP responded differently to the different methods of adding CIP to the soil.
Nitrogen-rich materials such as poultry litter (PL) contributes to substantial N and C loss in the form of ammonia (NH3) and carbon dioxide (CO2) during composting. Biochar can act as a sorbent of ammonia (NH3) and CO2 emission released during co-composting. Thus, co-composting poultry litter with rice husk biochar as a bulking agent is a good technique to mitigate NH3 volatilization and CO2 emission. A study was conducted to evaluate the effects of composting the mixtures of poultry litter with rice husk biochar at different ratios on NH3 and CO2 emissions. Four mixtures of poultry litter and rice husk biochar at different rate were composted at 0:1, 0.5:1, 1.3:1 and 2.3:1 ratio of rice husk biochar (RHB): poultry litter (PL) on a dry weight basis to achieve a suitable C/N ratio of 15, 20, 25, and 30, respectively. The results show that composting poultry litter with rice husk biochar can accelerate the breakdown of organic matter, thereby shortening the thermophilic phase compared to composting using poultry litter alone. There was a significant reduction in the cumulative NH3 emissions, which accounted for 78.38%, 94.60%, and 97.30%, for each C/N ratio of 20, 25, and 30. The total nitrogen (TN) retained relative was 75.96%, 85.61%, 90.24%, and 87.89% for each C/N ratio of 15, 20, 25, and 30 at the completion of composting. Total carbon dioxide lost was 5.64%, 6.62%, 8.91%, and 14.54%, for each C/N ratio of 15, 20, 21, and 30. In addition, the total carbon (TC) retained were 66.60%, 72.56%, 77.39%, and 85.29% for 15, 20, 25, and 30 C/N ratios and shows significant difference as compared with the initial reading of TC of the compost mixtures. In conclusion, mixing and composting rice husk biochar in poultry litter with C/N ratio of 25 helps in reducing the NH3 volatilization and CO2 emissions, while reducing the overall operational costs of waste disposal by shortening the composting time alongside nitrogen conservation and carbon sequestration. In formulating the compost mixture with rice husk biochar, the contribution of C and N from the biochar can be neglected in the determination of C/N ratio to predict the rate of mineralization in the compost because biochar has characteristic of being quite inert and recalcitrant in nature.
The livestock industry needs to use crop straws that are highly digestible to improve feed productivity and reduce ruminal methane emissions. Hence, this study aimed to use the ozonation and pelleting processes to enhance the digestibility and reduce the ruminal methane emissions of wheat straw enriched with two nitrogen sources (i.e., urea and heat-processed broiler litter). Various analyses were conducted on the pellets, including digestibility indicators, mechanical properties, surface chemistry functionalization, chemical-spectral-structural features, and energy requirements. For comparison, loose forms of the samples were also analyzed. The nitrogen-enriched ozonated wheat straw pellets had 43.06 % lower lignin, 28.30 % higher gas production for 24 h, 12.28 % higher metabolizable energy, 13.78 % higher in vitro organic matter digestibility for 24 h, and 28.81 % higher short-chain fatty acid content than the nitrogen-enriched loose sample. The reduction of methane emissions by rumen microorganisms of nitrogen-enriched wheat straw by ozonation, pelleting, and ozonation-pelleting totaled 89.15 %, 23.35 %, and 66.98 %, respectively. The ozonation process resulted in a 64 % increase in the particle density, a 5.5-time increase in the tensile strength, and a 75 % increase in the crushing energy of nitrogen-enriched wheat straw. In addition, ozone treatment could also reduce the specific and thermal energy consumption required in the pelleting process by 15.10 % and 7.61 %, respectively.
Stover and manure are the main solid waste in agricultural industry. The generation of stover and manure could lead to serious environmental pollution if not handled properly. Composting is the potential greener solution to remediate and reduce agricultural solid waste, through which stover and manure could be remediated and converted into organic fertilizer, but the long composting period and low efficiency of humic substance production are the key constraints in such remediation approach. In this study, we explore the effect of lignocellulose selective removal on composting by performing chemical pretreatment on agricultural waste followed by utilization of biochar to assist in the remediation by co-composting treatment and reveal the impacts of different lignocellulose component on organic fertilizer production. Aiming to discover the key factors that influence humification during composting process and improve the composting quality as well as comprehensive utilization of agricultural solid waste. The results demonstrated that the removal of selective lignin or hemicellulose led to the shift of abundances lignocellulose-degrading bacteria, which in turn accelerated the degradation of lignocellulose by almost 51.2%. The process also facilitated the remediation of organic waste via humification and increased the humic acid level and HA/FA ratio in just 22 days. The richness of media relies on their lignocellulose content, which is negatively correlated with total nitrogen content, humic acid (HA) content, germination index (GI), and pH, but positively correlated with fulvic acid (FA) and total organic carbon (TOC). The work provides a potential cost effective and efficient framework for agricultural solid waste remediation and reduction.
Contamination of rice by arsenic represents a significant human health risk. Roxarsone -bearing poultry manure is a major pollution source of arsenic to paddy soils. A mesocosm experiment plus a laboratory experiment was conducted to reveal the role of rainwater-borne H2O2 in the degradation of roxarsone in paddy rice soils. While roxarsone could be degraded via chemical oxidation by Fenton reaction-derived hydroxyl radical, microbially mediated decomposition was the major mechanism. The input of H2O2 into the paddy soils created a higher redox potential, which favored certain roxarsone-degrading and As(III)-oxidizing bacterial strains and disfavored certain As(V)-reducing bacterial strains. This was likely to be responsible for the enhanced roxarsone degradation and transformation of As(III) to As(V). Fenton-like reaction also tended to enhance the formation of Fe plaque on the root surface, which acted as a filter to retain As. The dominance of As(V) in porewater, combined with the filtering effect of Fe plaque significantly reduced the uptake of inorganic As by the rice plants and consequently its accumulation in the rice grains. The findings have implications for developing management strategies to minimize the negative impacts from the application of roxarsone-containing manure for fertilization of paddy rice soils.
Repeated applications of animal manure as fertilizer are normal agricultural practices that may release veterinary antibiotics and hormones into the environment from treated animals. Broiler manure samples and their respective manure-amended agricultural soil samples were collected in selected locations in the states of Selangor, Negeri Sembilan and Melaka in Malaysia to identify and quantify veterinary antibiotic and hormone residues in the environment. The samples were analyzed using ultrasonic extraction followed by solid phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The broiler manure samples were found to be contaminated with at least six target analytes, namely, doxycycline, enrofloxacin, flumequine, norfloxacin, trimethoprim and tylosin. These analytes were detected in broiler manure samples with maximum concentrations reaching up to 78,516 μg kg(-1) dry weight (DW) (doxycycline). For manure-amended agricultural soil samples, doxycycline and enrofloxacin residues were detected in every soil sample. The maximum concentration of antibiotic detected in soil was 1331 μg kg(-1) DW (flumequine). The occurrence of antibiotics and hormones in animal manure at high concentration poses a risk of contaminating agricultural soil via fertilization with animal manure. Some physico-chemical parameters such as pH, total organic carbon (TOC) and metal content played a considerable role in the fate of the target veterinary antibiotics and progesterone in the environment. It was suggested that these parameters can affect the adsorption of pharmaceuticals to solid environmental matrices.
Vermicomposting is commonly adopted for the treatment of livestock organic wastes. In the present study, two types of livestock manure were used for culturing of the earthworm, Eisenia foetida. Each treatment group consisted of six replicates and worm vermicasts were examined after 5 weeks. The concentrations of total C, P and K in goat manure vermicasts were higher than those in cattle manure vermicasts. Cattle vermicasts had a higher N content than goat vermicasts but the C:N ratio of fresh manure was higher than that of vermicasts for both materials. Earthworm biomass and reproductive performance, in terms of number of worms after 5 weeks of experiment, were higher in cattle manure than in goat manure. The cocoon production per worm in cattle manure was higher than in goat manure. However, the hatchability of cocoons was not affected by manure treatments. In conclusion, cattle manure provided a more nutritious and friendly environment to the earthworms than goat manure.
The behavior of veterinary antibiotics in the soil is commonly studied using the following methods to add antibiotics to the soil: (A) adding manure collected from animals fed a diet that includes antibiotics; (B) adding antibiotic-free animal manure spiked with antibiotics; and (C) the direct addition of antibiotics. However, most studies have only used methods (B) and (C) in their research, and few studies have simultaneously compared the different antibiotic addition methods. This study used tylosin A (TYLA) as a model antibiotic to compare the effects of these three commonly used antibiotic addition methods on the dissipation rates of TYLA and the numbers of resistance genes in laboratory incubation experiments. The results showed that the three treatment methods produced similar TYLA degradation trends; however, there were significant differences (P<0.05) in the TYLA degradation half-life (t1/2) among the three methods. The half-life of TYLA degradation in treatments A, B and C was 2.44 ± 0.04, 1.21 ± 0.03 and 5.13 ± 0.11 days, respectively. The presence of manure resulted in a higher electrical conductivity (EC), higher relative abundance of Citrobacter amalonaticus, higher macrolide resistant gene (ermB, ermF and ermT) count and lower ecological toxicity in the soil, which could partially explain the higher TYLA degradation rate in the treatments containing manure. The higher degradation rate of TYLA in treatment B when compared to treatment A could be due to the lower concentrations of tylosin B (TYLB) and tylosin D (TYLD). The main route for veterinary antibiotics to enter the soil is via the manure of animals that have been administered antibiotics. Therefore, the more appropriate method to study the degradation and ecotoxicity of antibiotic residues in the soil is by using manure from animals fed/administered the particular antibiotic rather than by adding the antibiotic directly to the soil.
The availability of paddy husk from rice processing plants remains high owing to increase in the worldwide rice consumption. Increasing demand for chicken products leads to poultry wastes production. Co-composting of the aforementioned wastes could solve the indiscriminate disposal of these wastes. Thus, co-composting of paddy husk and chicken slurry with clinoptilolite zeolite and urea as additive was carried out. Clinoptilolite zeolite was used to enhance ammonium and nitrate retention in the compost. Temperature of the compost was monitored three times daily for 55 days. Cation exchange capacity, organic matter, ash, humic acids, pH, total C, N, C/N ratio; total P, exchangeable Ca, Mg, K, NH4+, NO3-, and heavy metals contents were determined using standard procedures. pH, total N, humic acids, ash, NH4+, NO3-, P, Ca, Mg, and K contents increased but the salinity, heavy metals contents, and microbial population were low after the co-composting process. Zea mays L. (test crop) seed germination rate in distilled water and the compost were not significantly different. Growth of Spinach oleracea (test crop) on a peat-based growing medium and the compost was also not significantly different. These findings were possible because the clinoptilolite zeolite used in co-composting reduced accumulation of heavy metals that may have damage effects on the test crops. Mature compost with good agronomic properties can be produced by co-composting chicken slurry and paddy husk using clinoptilolite zeolite and urea as additives.
Ammonia emissions is an important issue during composting because it can cause secondary pollution and a significant of nitrogen loss. Based on research adding Bacillus stearothermophilus can reduce ammonia emissions during composting because it can use sugar in organic matter fermentation to produce organic acids over 50 °C. This study conducted the batch experiments by adding different concentrations of Bacillus stearothermophilus to reduce the ammonia emissions and find out its characteristic during layer manure composting by using an aerobic composting reactor with sawdust as a bulking agent. The results show that the application of Bacillus stearothermophilus can accelerate the rate of temperature and significantly decrease pH, the warming period was 2 days in the treatment with Bacillus stearothermophilus, while it was 4 days in the treatment without Bacillus stearothermophilus. Ammonia emissions were mainly occurred in warming and high temperature period during composting. The ammonia emissions in the treatment with 8.00 g/kg initial Bacillus stearothermophilus were significantly lower than the other lower Bacillus stearothermophilus treatment and control during composting (p 0.05). MiSeq System Sequencing results find that the addition of Bacillus stearothermophilus changed the bacterial community structure under warming and high-temperature periods during composting, increased the relative abundance of lactic acid bacillus and nitrification bacteria. Therefore, the reason for the low ammonia emission in 8.00 g/kg initial Bacillus stearothermophilus treatments might be not only due to the Bacillus stearothermophilus itself, but also Bacillus stearothermophilus can change the indigenous microorganism community, including increase the relative content of lactic acid Bacillus and nitrification bacteria, thus reducing the pH and promoting nitrification, and reducing ammonia emissions.
This study's primary purpose was to investigate the possible amelioration of limited irrigation conditions by mycorrhiza (AMF), vermicompost, and green manure for lingrain plants. This experiment was accomplished as a factorial based on the completely randomized design with three replications. The first factor was green manure (without green manure and with Trifolium pratense as green manure); the second factor consisted of Rhizophagus irregularis mycorrhiza, vermicompost, a combination of mycorrhiza and vermicompost and none of them, and also the third factor was irrigation regime (full irrigation and late-season water limitation). Green manure, vermicompost, and mycorrhiza single-use enhanced the plant's underwater limitation conditions compared to the control. However, vermicompost and green manure or mycorrhiza developed a positive synergistic effect on most traits. Combining green manure with the dual fertilizer (mycorrhiza + vermicompost) resulted in the vermicompost and mycorrhiza synergistic effects, especially under limited irrigation. Consequently, the combination of green manure, mycorrhiza, and vermicompost experienced the highest amount of leaf relative water content, root colonization, leaf nitrogen, chlorophyll a, chlorophyll b, carotenoids, antioxidant enzymes activity, grain yield, and oil yield, which would lead to more resistance of plants to limited irrigation conditions.
Sago bark (SB) and empty fruit bunch (EFB) are available abundantly as agricultural waste in Sarawak. This study was conducted to investigate the physicochemical characteristics of SB and EFB as composting materials and used as a plant growth medium. The SB and EFB composts were prepared in a separate container by mixing chicken manure as compost accelerator and wood chips as a bulking agent in dry weight equivalent ratio (1:1:1). The maturity and stability of compost in 60-day composting periods were evaluated via physicochemical characterization of the composts in terms of pH, elemental content, total ash content, moisture content and nutrient analyses. The effect of the compost usage as growth medium was assessed towards water spinach and green mustard via seed germination and pot study. After 2 months, the colour of both composts was dark brown with an earthy smell. The acidic pH of the initial composting stage has changed into alkaline pH after 60 days of composting. Total NPK present in the SB and EFB composts were 0.96% and 1.21%, respectively. The germination index (GI) for the studied vegetables was above 100%, while the pot study showed that vegetables in compost media has higher growth compared to the control, after 14 days. SB and EFB are renewable waste which can be used as an excellent compost and able to improve the quality of the soil.
Vermicomposting is a process in which earthworms are used to convert organic materials into humus-like material known as vermicompost. A number of researchers throughout the world have found that the nutrient profile in vermicompost is generally higher than traditional compost. In fact, vermicompost can enhance soil fertility physically, chemically and biologically. Physically, vermicompost-treated soil has better aeration, porosity, bulk density and water retention. Chemical properties such as pH, electrical conductivity and organic matter content are also improved for better crop yield. Nevertheless, enhanced plant growth could not be satisfactorily explained by improvements in the nutrient content of the soil, which means that other plant growth-influencing materials are available in vermicomposts. Although vermicomposts have been shown to improve plant growth significantly, the application of vermicomposts at high concentrations could impede growth due to the high concentrations of soluble salts available in vermicomposts. Therefore, vermicomposts should be applied at moderate concentrations in order to obtain maximum plant yield. This review paper discusses in detail the effects of vermicompost on soil fertility physically, chemically and biologically. Future prospects and economy on the use of organic fertilizers in the agricultural sector are also examined.
The excretion rates and ecological risk to the environment of three commonly used veterinary antibiotics (VAs), amoxicillin, ciprofloxacin, and doxycycline, in layer hen manure during the application and withdrawal periods were investigated in a study consisting of a control group fed with VA-free basal diet and nine treatment groups consisted of three levels (200 mg/kg, 100 mg/kg, and 50 mg/kg) of amoxicillin (AMX), ciprofloxacin (CIP), or doxycycline (DOC). Each treatment group was replicated seven times with three layer hens per replication. Results of the study showed that the average excretion rates of AMX in the 200, 100, and 50 mg/kg groups were 67.88, 55.82, and 66.15%, respectively, while those for CIP and DOC were 47.84, 51.85, and 44.87% and 82.67, 94.39, and 95.72%, respectively. The concentrations of the above veterinary drugs in manure decreased sharply in the withdrawal period (7, 28, and 10 d, respectively), for AMX, DOC, and CIP. Neither AMX nor DOC was detected in the manure after the withdrawal period. In contrast to AMX and DOC, the excretion rate of CIP was significantly lower and thus had a longer residence time. Ecological risk study, estimated using hazard quotient values, showed that AMX in the 100 and 50 mg/kg groups posed no risk to the environment after d 1 of withdrawal, while CIP in the 50 mg/kg group posed no risk to the environment from d 5 of withdrawal. CIP in the 200 and 100 mg/kg groups required 10 d withdrawal in order to pose no risk to the environment. In contrast, DOC residue during withdrawal in the manure posed no risk to the environment, thus making it more environmentally safe.
Cat manure (CM) possesses high level of nutrients for growing food crop. However, animal manure may contain toxic elements that may contaminate food crop. Spent coffee ground (SCG) may be used to reduce mobility of heavy metals and reduce crop uptake. In this study, SCG was composted with CM for 31 days to produce a co-compost (SCG-CM) for growing spinach (Spinacia oleracea). The growth rate of spinach was assessed until its maturity, and the metal uptake of spinach shoot was determined thereafter using inductively coupled plasma-optical emission spectroscopy (ICP-OES). The effect of soil treatment with SCG-CM on the height and elemental composition of spinach were compared with that of chicken manure compost (CMC). The prepared composts were primarily organic matter (72.9-81.4 % w/w) with the rest are ash (13.3-23.4 % w/w) and moisture (1.2-2.6 % w/w). Zinc content in SCG-CM (1261 ± 0.1 mg/kg) is significantly higher than that of soil and CMC (p < 0.05) and has exceeded the maximum permissible limit set by European Union Standard (2002) and the Malaysian Compost Quality Standard and Guidelines (2000). Matured spinach reached maximum plant height after 33 days. The amendment of SCG-CM significantly increased the height of spinach (32 ± 6 cm) compared to that of CMC (13 ± 1 cm) (p < 0.05). However, contents of Zn, Cu, Pb and Cd were not increased for spinach grown in the SCG-CM-amended soil, and the level of those elements are below permissible limit set by the Malaysian Food Act 1983 and Food Regulations 1985. This study shows that SCG-CM is effective in improving yield without causing accumulation of toxic trace elements in spinach.
In this study, the differences on the physico-chemical parameters, lignocellulose degradation, dynamic succession of microbial community, gene expression of carbohydrate-active enzymes and antibiotics resistance genes were compared during composting systems of bagasse pith/pig manure (BP) and manioc waste/pig manure (MW). The results revealed that biodegradation rates of organic matter, cellulose, hemicellulose and lignin (29.14%, 17.53%,45.36% and 36.48%) in BP were higher than those (15.59%, 16.74%, 41.23% and 29.77%) in MW. In addition, the relative abundance of Bacillus, Luteimonas, Clostridium, Pseudomonas, Streptomyces and expression of genes encoding carbohydrate- active enzymes in BP were higher than those in MW based on metagenomics sequencing. During composting, antibiotics and antibiotic resistance genes were substantially reduced, but the removal efficiency was divergent in the both samples. Taken together, metagenomics analysis was a potential method for evaluating lignocellulose's biodegradation process and determining the elimination of antibiotic and antibiotic resistance genes from different composting sources of biomass.
As the population increases, energy demands continue to rise rapidly. In order to satisfy this increasing energy demand, biogas offers a potential alternative. Biogas is economically viable to be produced through anaerobic digestion (AD) from various biomass feedstocks that are readily available in Malaysia, such as food waste (FW), palm oil mill effluent (POME), garden waste (GW), landfill, sewage sludge (SS) and animal manure. This paper aims to determine the potential feedstocks for biogas production via AD based on their characteristics, methane yield, kinetic studies and economic analysis. POME and FW show the highest methane yield with biogas yields up to 0.50 L/g VS while the lowest is 0.12 L/g VS by landfill leachate. Kinetic study shows that modified Gompertz model fits most of the feedstock with R 2 up to 1 indicating that this model can be used for estimating treatment efficiencies of full-scale reactors and performing scale-up analysis. The economic analysis shows that POME has the shortest payback period (PBP), highest internal rate of return (IRR) and net present value (NPV). However, it has already been well explored, with 93% of biogas plants in Malaysia using POME as feedstock. The FW generation rate in Malaysia is approximately 15,000 tonnes per day, at the same time FW as the second place shows potential to have a PBP of 5.4 years and 13.3% IRR, which is close to the results achieved with POME. This makes FW suitable to be used as the feedstock for biogas production.
Lignocellulosic biomasses such as wheat straw are widely used as a feedstock for biogas production. However, these biomasses are mainly composed of a compact fibre structure and therefore, it is recommended to treat them prior to its usage for biogas production in order to improve their bioavailability. The aim of this work is to evaluate, in terms of performance stability, methane yield and economic feasibility, two different scenarios: a mesophilic codigestion of wheat straw and animal manure with or without a low-energy demand alkaline pre-treatment (0.08gKOHgTS-1of wheat straw, for 24h and at 25°C). Besides this, said pre-treatment was also analysed based on the improvement of the bioavailable carbohydrate content in the untreated versus the pre-treated wheat straw. The results pointed out that pre-treated wheat straw prompted a more stable performance (in terms of pH and alkalinity) and an improved methane yield (128% increment) of the mesophilic codigestion process, in comparison to the "untreated" scenario. The pre-treatment increased the content of cellulose, hemicellulose and other compounds (waxes, pectin, oil, etc.) in the liquid fraction, from 5% to 60%, from 11.5% to 39.1% TS and from 57% to 79% of the TS in the liquid fraction for the untreated and pre-treated wheat straws, respectively. Finally, the pre-treated scenario gained an energy surplus of a factor 13.5 and achieved a positive net benefit of 90.4€tVS-WS-1d-1, being a favourable case for an eventual scale-up of the combined process.