Nitrogen (N) is a macronutrient desired by crop plants in large quantities. However, hiking fertilizer prices need alternative N sources for reducing its requirements through appropriate management practices. Plant growth promoting rhizobacteria (PGPR) are well-known for their role in lowering N requirements of crop plants. This study assessed the impact of PGPR inoculation on growth, allometry and biochemical traits of chili under different N doses. Two PGPR, i.e., Azospirillum 'Er-20' (nitrogen fixing) and Agrobacterium 'Ca-18' (phosphorous solubilizing) were used for inoculation, while control treatment had no PGPR inoculation. Six N doses, i.e., 100, 80, 75, 70, 60 and 50% of the N required by chili were included in the study. Data relating to growth traits, biochemical attributes and yield related traits were recorded. Interaction among N doses and PGPR inoculation significantly altered all growth traits, biochemical attributes and yield related traits. The highest values of the recorded traits were observed for 100% N with and without PGPR inoculation and 75% N with PGPR inoculation. The lowest values of the recorded traits were noted for 50% N without PGPR inoculation. The PGPR inoculation improved the measured traits compared to the traits recorded noted in same N dose without PGPR inoculation. Results revealed that PGPR had the potential to lower 25% N requirement for chili. Therefore, it is recommended that PGPR must be used in chili cultivation to lower N requirements.
Nitrate-nitrogen leaching from agricultural areas is a major cause for groundwater pollution. Polluted groundwater with high levels of nitrate is hazardous and cause adverse health effects. Human consumption of water with elevated levels of NO3-N has been linked to the infant disorder methemoglobinemia and also to non-Hodgkin's disease lymphoma in adults. This research aims to study the temporal patterns and source apportionment of nitrate-nitrogen leaching in a paddy soil at Ladang Merdeka Ismail Mulong in Kelantan, Malaysia. The complex data matrix (128 x 16) of nitrate-nitrogen parameters was subjected to multivariate analysis mainly Principal Component Analysis (PCA) and Discriminant Analysis (DA). PCA extracted four principal components from this data set which explained 86.4% of the total variance. The most important contributors were soil physical properties confirmed using Alyuda Forecaster software (R2 = 0.98). Discriminant analysis was used to evaluate the temporal variation in soil nitrate-nitrogen on leaching process. Discriminant analysis gave four parameters (hydraulic head, evapotranspiration, rainfall and temperature) contributing more than 98% correct assignments in temporal analysis. DA allowed reduction in dimensionality of the large data set which defines the four operating parameters most efficient and economical to be monitored for temporal variations. This knowledge is important so as to protect the precious groundwater from contamination with nitrate.
Bioretention systems are among the most popular stormwater best management practices (BMPs) for urban runoff treatment. Studies on plant performance using bioretention systems have been conducted, especially in developed countries with a temperate climate, such as the USA and Australia. However, these results might not be applicable in developing countries with tropical climates due to the different rainfall regimes and the strength of runoff pollutants. Thus, this study focuses on the performance of tropical plants in treating urban runoff polluted with greywater using a bioretention system. Ten different tropical plant species were triplicated and planted in 30 mesocosms with two control mesocosms without vegetation. One-way ANOVA was used to analyze the performance of plants, which were then ranked based on their performance in removing pollutants using the total score obtained for each water quality test. Results showed that vetiver topped the table with 86.4% of total nitrogen (TN) removal, 93.5% of total phosphorus (TP) removal, 89.8% of biological oxygen demand (BOD) removal, 90% of total suspended solids (TSS) removal, and 92.5% of chemical oxygen demand (COD) removal followed by blue porterweed, Hibiscus, golden trumpet, and tall sedge which can be recommended to be employed in future bioretention studies.
Bioretention systems have been implemented as stormwater best management practices (BMPs) worldwide to treat non-point sources pollution. Due to insufficient research, the design guidelines for bioretention systems in tropical countries are modeled after those of temperate countries. However, climatic factors and stormwater runoff characteristics are the two key factors affecting the capacity of bioretention system. This paper reviews and compares the stormwater runoff characteristics, bioretention components, pollutant removal requirements, and applications of bioretention systems in temperate and tropical countries. Suggestions are given for bioretention components in the tropics, including elimination of mulch layer and submerged zone. More research is required to identify suitable additives for filter media, study tropical shrubs application while avoiding using grass and sedges, explore function of soil faunas, and adopt final discharged pollutants concentration (mg/L) on top of percentage removal (%) in bioretention design guidelines.
This work studied the bioremediation of weathered crude oil (WCO) in coastal sediment samples using central composite face centered design (CCFD) under response surface methodology (RSM). Initial oil concentration, biomass, nitrogen and phosphorus concentrations were used as independent variables (factors) and oil removal as dependent variable (response) in a 60 days trial. A statistically significant model for WCO removal was obtained. The coefficient of determination (R(2)=0.9732) and probability value (P<0.0001) demonstrated significance for the regression model. Numerical optimization based on desirability function were carried out for initial oil concentration of 2, 16 and 30 g per kg sediment and 83.13, 78.06 and 69.92 per cent removal were observed respectively, compare to 77.13, 74.17 and 69.87 per cent removal for un-optimized results.
The characteristics of urban stormwater pollution in the tropics are still poorly understood. This issue is crucial to the tropical environment because its rainfall and runoff generation processes are so different from temperate regions. In this regard, a stormwater monitoring program was carried out at three urban catchments (e.g. residential, commercial and industrial) in the southern part of Peninsular Malaysia. A total of 51 storm events were collected at these three catchments. Samples were analyzed for total suspended solids, 5-day biochemical oxygen demand, chemical oxygen demand (COD), oil and grease, nitrate nitrogen, nitrite nitrogen, ammonia nitrogen (NH3-N), soluble reactive phosphorus and total phosphorus. Principal component analysis (PCA) and hierarchical cluster analysis were used to interpret the stormwater quality data for pattern recognition and identification of possible sources. The most likely sources of stormwater pollutants at the residential catchment were from surface soil and leachate of fertilizer from domestic lawns and gardens, whereas the most likely sources for the commercial catchment were from discharges of food waste and washing detergent. In the industrial catchment, the major sources of pollutants were discharges from workshops and factories. The PCA factors further revealed that COD and NH3-N were the major pollutants influencing the runoff quality in all three catchments.
Water hyacinth (Eichhornia crassipes) and water lettuce (Pistia stratiotes) were analyzed to determine their effectiveness in aquaculture wastewater treatment in Malaysia. Wastewater from fish farm in Semanggol Perak, Malaysia was sampled and the parameters determined included, the pH, turbidity, dissolved oxygen (DO), chemical oxygen demand (COD), biochemical oxygen demand (BOD), nitrite phosphate (PO4(3-)), nitrate (NO(3-)), nitrite (NO(-2)), ammonia (NH3), and total kjedahl nitrogen (TKN). Also, hydroponics system was set up and was added with fresh plants weights of 150 +/- 20 grams Eichhornia crassipes and 50 +/- 10 grams Pistia stratiotes during the 30 days experiment. The phytoremediation treatment with Eichhornia crassipes had pH ranging from 5.52 to 5.59 and from 4.45 to 5.5 while Pistia stratiotes had its pH value from 5.76 to 6.49 and from 6.24 to 7.07. Considerable percentage reduction was observed in all the parameters treated with the phytoremediators. Percentage reduction of turbidity for Eichhornia crassipes were 85.26% and 87.05% while Pistia stratiotes were 92.70% and 93.69% respectively. Similar reductions were observed in COD, TKN, NO(3-), NH3, and PO4(3-). The capability of these plants in removing nutrients was established from the study. Removal of aquatic macrophytes from water bodies is recommended for efficient water purification.
Vegetation restoration is a widely used, effective, and sustainable method to improve soil quality in post-mining lands. Here we aimed to assess global patterns and driving factors of potential vegetation restoration effects on soil carbon, nutrients, and enzymatic activities. We synthesized 4838 paired observations extracted from 175 publications to evaluate the effects that vegetation restoration might have on the concentrations of soil carbon, nitrogen, and phosphorus, as well as enzymatic activities. We found that (1) vegetation restoration had consistent positive effects on the concentrations of soil organic carbon, total nitrogen, available nitrogen, ammonia, nitrate, total phosphorus, and available phosphorus on average by 85.4, 70.3, 75.7, 54.6, 58.6, 34.7, and 60.4 %, respectively. Restoration also increased the activities of catalase, alkaline phosphatase, sucrase, and urease by 63.3, 104.8, 125.5, and 124.6 %, respectively; (2) restoration effects did not vary among different vegetation types (i.e., grass, tree, shrub and their combinations) or leaf type (broadleaved, coniferous, and mixed), but were affected by mine type; and (3) latitude, climate, vegetation species richness, restoration year, and initial soil properties are important moderator variables, but their effects varied among different soil variables. Our global scale study shows how vegetation restoration can improve soil quality in post-mining lands by increasing soil carbon, nutrients, and enzymatic activities. This information is crucial to better understand the role of vegetation cover in promoting the ecological restoration of degraded mining lands.
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 nitrogen and nitrate-nitrogen concentration, reduce pH (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.
The effect of metals on environmental health is well documented and monitoring these and other pollutants is considered an important part of environmental management. Developing countries are yet to fully appreciate the direct impacts of pollution on aquatic ecosystems and as such, information on pollution dynamics is scant. Here, we assessed the temporal and spatial dynamics of stream sediment metal and nutrient concentrations using contaminant indices (e.g. enrichment factors, pollution load and toxic risk indices) in an arid temperate environment over the wet and dry seasons. The mean sediment nutrient, organic matter and metal concentration were highest during the dry season, with high values being observed for the urban environment. Sediment contaminant assessment scores indicated that during the wet season, the sediment quality was acceptable, but not so during the dry season. The dry season had low to moderate levels of enrichment for metals B, Cu, Cr, Fe, Mg, K and Zn. Overall, applying the sediment pollution load index highlighted poor quality river sediment along the length of the river. Toxic risk index indicated that most sites posed no toxic risk. The results of this study highlighted that river discharge plays a major role in structuring temporal differences in sediment quality. It was also evident that infrastructure degradation was likely contributing to the observed state of the river quality. The study contributes to our understanding of pollution dynamics in arid temperate landscapes where vast temporal differences in base flow characterise the riverscape. Such information is further useful for contrasting sediment pollution dynamics in aquatic environments with other climatic regions.
The fluctuation of domestic wastewater characteristic inhibits the current conventional microbial-based treatment. The bioremediation fungi has received attention and reported to be an effective alternative to treat industrial wastewater. Similar efficient performance is envisaged for domestic wastewater whereby assessed performance of fungi for varying carbon-to-nitrogen ratios in domestic wastewater is crucial. Thus, the performance of pre-grown wild-Serbian Ganoderma lucidum mycelial pellets (GLMPs) was evaluated on four different synthetic domestic wastewaters under different conditions of initial pH (pH 4, 5, and 7) and chemical oxygen demand (COD) to nitrogen (COD/N) ratio of 3.6:1, 7.1:1, 14.2:1, and 17.8:1 (C3.6N1, C7.1N1, C14.2N1, and C17.8N1). The COD/N ratios with a constant concentration of ammonia-nitrogen (NH3-N) were chosen on the basis of the urban domestic wastewater characteristics sampled at the inlet basin of a sewage treatment plant (STP). The parameters of pH, COD, and NH3-N were measured periodically during the experiment. The wild-Serbian GLMPs efficiently removed the pollutants from the synthetic sewage. The COD/N ratio of C17.8N1 wastewater had the best COD and NH3-N removal, as compared to the lower COD/N ratio, and the shortest treatment time was obtained in an acidic environment at pH 4. The highest percentage for COD and NH3-N removal achieved was 96.0% and 93.2%, respectively. The results proved that the mycelium of GLMP has high potential in treating domestic wastewater, particularly at high organic content as a naturally sustainable bioremediation system.
The tropical estuarine ecosystem is fascinating for studying the dynamics of water quality and phytoplankton diversity due to its frequently changing hydrological conditions. Most importantly, phytoplankton is the main supplier of ω3 polyunsaturated fatty acids (PUFA) in the coastal food web for fish as they could not synthesize PUFA. This study evaluated seasonal variations of water quality parameters in the Meghna River estuary (MRE), explored how phytoplankton diversity changes according to hydro-chemical parameters, and identified the major phytoplankton groups as the main source of PUFA for hilsa fish. Ten water quality indicators including temperature, dissolved oxygen, pH, salinity, dissolved inorganic nitrogen (DIN = nitrate, nitrite, ammonia) and phosphorus, dissolved silica and chlorophyll-a were evaluated. In addition, phytoplankton diversity was assessed in the water and hilsa fish gut. Principal component analysis (PCA) was used to analyze the spatio-temporal changes in the water quality conditions, and the driving factors in the MRE. Four main components were extracted and explained 75.4% variability of water quality parameters. The most relevant driving factors were dissolved oxygen, salinity, temperature, and DIN (nitrate, nitrite and ammonia). These variabilities in physicochemical parameters and dissolved inorganic nutrients caused seasonal variations in two major groups of phytoplankton. Peak abundance of Chlorophyta (green algae) occurred in water in nutrient-rich environments (nitrogen and phosphorus) during the wet (36%) season, while Bacillariophyta (diatoms) were dominant during the dry (32%) season that depleted dissolved silica. Thus, the decrease of green algae and the increase of diatoms in the dry season indicated the potential link to seasonal changes of hydro-chemical parameters. The green algae (53.7%) were the dominant phytoplankton group in the hilsa gut content followed by diatoms (22.6%) and both are contributing as the major source of PUFAs for hilsa fish according to the electivity index as they contain the highest amounts of PUFAs (60 and 28% respectively).
This study analyzes and compares the results of leachate composition at the semi-aerobic Pulau Burung Landfill Site (PBLS) (unaerated pond and intermittently aerated pond) and the anaerobic Kulim Sanitary Landfill in the northern region of Malaysia. The raw samples were collected and analyzed for twenty parameters. The average values of the parameters such as phenols (1.2, 6.7, and 2.6 mg/L), total nitrogen (448, 1200, and 300 mg/L N-TN), ammonia-N (542, 1568, and 538 mg/L NH(3)-N), nitrite (91, 49, and 52 mg/L NO(2)(-)-N), total phosphorus (21, 17, and 19 mg/L), BOD(5) (83, 243, and 326 mg/L), COD (935, 2345, and 1892 mg/L), BOD(5)/COD (0.096,0.1124,0.205%), pH (8.20, 8.28, and 7.76), turbidity (1546, 180, and 1936 Formazin attenuation units (FAU)), and color (3334, 3347, and 4041 Pt Co) for leachate at the semi-aerobic PBLS (unaerated and intermittently aerated) and the anaerobic Kulim Sanitary Landfill were recorded, respectively. The obtained results were compared with previously published data and data from the Malaysia Environmental Quality Act 1974. The results indicated that Pulau Burung leachate was more stabilized compared with Kulim leachate. Furthermore, the aeration process in PBLS has a considerable effect on reducing the concentration of several pollutants. The studied leachate requires treatment to minimize the pollutants to an acceptable level prior to discharge into water courses.
Acid sulfate, peat, sandy podzolic, and saline soils are widely distributed in the lowlands of Thailand and Malaysia. The nutrient concentrations in the leaves of plants grown in these type of soils were studied with the aim of developing a nutritional strategy for adapting to such problem soils. In sago and oil palms that were well-adapted to peat soil, the N, P, and K concentrations were the same in the mature leaves, while the Ca, Mg, Na, and Fe concentrations were higher in the mature leaves of the oil palm than of the sago palm. Melastoma malabathricum and Melaleuca cajuputi plants that were well-adapted to low pH soils, peat. and acid sulfate soils were also studied. It was observed that a high amount of Al accumulated in the M. marabathricum leaves, while Al did not accumulate in M. cajuputi leaves. M. cajuputi plants accumulated large amounts of Na in their leaves or stems regardless of the exchangeable Na concentration in the soil, while M. malabathricum that was growing in saline-affected soils excluded Na. Positive relationships between macronutrients were recognized between P and N, between K and N, and between P and K. Al showed antagonistic relationships with P, K, Ca, Mg, Fe, Zn, Cu, and Na. Na also showed antagonistic relationships with P, K, Zn, Mn, Cu, and Al. Fe showed weak antagonistic relationships with Zn, Mn, Cu, and Al.
Nitrogen (N) transport from land to water is a dominant contributor of N in estuarine waters leading to eutrophication, harmful algal blooms, and hypoxia. Our objectives were to (1) investigate the composition of inorganic and organic N forms, (2) distinguish the sources and biogeochemical mechanisms of nitrate-N (NO3-N) transport using stable isotopes of NO3- and Bayesian mixing model, and (3) determine the dissolved organic N (DON) bioavailability using bioassays in a longitudinal gradient from freshwater to estuarine ecosystem located in the Tampa Bay, Florida, United States. We found that DON was the most dominant N form (mean: 64%, range: 46-83%) followed by particulate organic N (PON, mean: 22%, range: 14-37%), whereas inorganic N forms (NOx-N: 7%, NH4-N: 7%) were 14% of total N in freshwater and estuarine waters. Stable isotope data of NO3- revealed that nitrification was the main contributor (36.4%), followed by soil and organic N sources (25.5%), NO3- fertilizers (22.4%), and NH4+ fertilizers (15.7%). Bioassays showed that 14 to 65% of DON concentrations decreased after 5-days of incubation indicating utilization of DON by microbes in freshwater and estuarine waters. These results suggest that despite low proportion of inorganic N forms, the higher concentrations and bioavailability of DON can be a potential source of N for algae and bacteria leading to water quality degradation in the estuarine waters.
Stormwater runoff is recognized as a cause of water quality degradation because it may carry nitrogen (N) and other pollutants to aquatic ecosystems. Stormwater ponds are a stormwater control measure often used to manage stormwater runoff by holding a permanent pool of water, which reduces the peak flow, magnitude of runoff volume, and concentrations of nutrients and pollutants. We instrumented the outlet of a stormwater pond in an urban residential neighbourhood in Florida, United States to (1) investigate the concentration and composition of N forms during the summer rainy season (May to September 2016), and (2) determine the bioavailability of organic N in the stormwater pond with a bioassay experiment. A total of 144 outflow water samples over 13 storm events were collected at the outlet of the stormwater pond that collects runoff from the residential catchment. Samples were analysed for various inorganic N [ammonium (NH4-N), nitrate (NO3-N)], and organic N forms [dissolved organic nitrogen (DON), and particulate organic nitrogen (PON)]. Flow-weighted mean concentration of total N (TN) in pond outflow for all collected storm events was 1.3±1.42 mg L-1, with DON as the dominant form (78%), followed by PON and NO3-N (each at 8%), and NH4-N (6%). In the bioassay experiment, organic N (DON+PON) was significantly decreased by 25-28% after 5 days of incubation, suggesting that a portion of the DON carried from the pond outflow to receiving water bodies may be bioavailable. These results suggest that efforts to mitigate stormwater N outflows from urban ponds should incorporate both inorganic and organic N in management plans.
The impact of floating net cages culturing the seabass, Lates calcarifer, on planktonic processes and water chemistry in two heavily used mangrove estuaries in Malaysia was examined. Concentrations of dissolved inorganic and particulate nutrients were usually greater in cage vs. adjacent (approximately 100 m) non-cage waters, although most variability in water-column chemistry related to water depth and tides. There were few consistent differences in plankton abundance, production or respiration between cage and non-cage sites. Rates of primary production were low compared with rates of pelagic mineralization reflecting high suspended loads coupled with large inputs of organic matter from mangrove forests, fishing villages, fish cages, pig farms and other industries within the catchment. Our preliminary sampling did not reveal any large-scale eutrophication due to the cages. A crude estimate of the contribution of fish cage inputs to the estuaries shows that fish cages contribute only approximately 2% of C but greater percentages of N (32-36%) and P (83-99%) to these waters relative to phytoplankton and mangrove inputs. Isolating and detecting impacts of cage culture in such heavily used waterways--a situation typical of most mangrove estuaries in Southeast Asia--are constrained by a background of large, highly variable fluxes of organic material derived from extensive mangrove forests and other human activities.
Non-point source (NPS) pollution has become a vital contaminant source affecting the water environment because of its wide distribution, hydrodynamic complexity, and difficulty in prevention and control. In this study, the identification and evaluation of NPS pollution risk based on landscape pattern were carried out in the Hanjiang River basin above Ankang hydrological section, Shaanxi province, China. Landscape distribution information was obtained through land use data, analyzing the contribution of "source-sink" landscape to NPS pollution through the location-weighted landscape contrast index. Using the NPS pollution risk index to identify and evaluate the regional NPS pollution risk considering the slope, cost distance, soil erosion, and precipitation erosion affect migration of pollutants. The results showed that (i) the pollution risk was generally high in the whole watershed, and the sub-watersheds dominated by "source" landscapes account for 74.61% of the whole basin; (ii) the high-risk areas were distributed in the central, eastern, and western regions of the river basin; the extremely high-risk areas accounted for 12.7% of the whole watershed; and the southern and northern regions were dominated by forestland and grassland with little pollution risk; (iii) "source" landscapes were mostly distributed in areas close to the river course, which had a great impact on environment, and the landscape pattern units near the water body needed to be further adjusted to reduce the influence of NPS pollution.
In this study, the treatment of septage (originating from septic tanks) was carried out in a pilot-scale, two-staged, vertical-flow engineered wetland (VFEW). Palm kernel shells (PKS) were incorporated as part of the VFEW's substrate (B-PKS), to compare its organic matter (OM) and nitrogen (N) removal efficiency against wetlands with only sand substrates (B-SD). The results revealed satisfactory OM removal with >90% reduction efficiencies at both wetlands B-PKS and B-SD. No increment of chemical oxygen demand (COD) concentration was observed in the effluent of B-PKS. Ammonia load removal efficiencies were comparable (>91% and 95% in wetland B-PKS and B-SD, respectively). However, nitrate accumulation was observed in the effluent of B-SD where PKS was absent. This was due to the limited denitrification in B-SD, as sand is free of carbon. A lower nitrate concentration was associated with higher COD concentration in the effluent at B-PKS. This study has shown that the use of PKS was effective in improving the N removal efficiency in engineered wetlands.
Understanding the main driving factors of oasis river nutrients in arid areas is important to identify the sources of water pollution and protect water resources. Twenty-seven sub-watersheds were selected in the lower oasis irrigated agricultural reaches of the Kaidu River watershed in arid Northwest China, divided into the site, riparian, and catchment buffer zones. Data on four sets of explanatory variables (topographic, soil, meteorological elements, and land use types) were collected. The relationships between explanatory variables and response variables (total phosphorus, TP and total nitrogen, TN) were analyzed by redundancy analysis (RDA). Partial least squares structural equation modeling (PLS-SEM) was used to quantify the relationship between explanatory as well as response variables and fit the path relationship among factors. The results showed that there were significant differences in the TP and TN concentrations at each sampling point. The catchment buffer exhibited the best explanatory power of the relationship between explanatory and response variables based on PLS-SEM. The effects of various land use types, meteorological elements (ME), soil, and topography in the catchment buffer were responsible for 54.3% of TP changes and for 68.5% of TN changes. Land use types, ME and soil were the main factors driving TP and TN changes, accounting for 95.56% and 94.84% of the total effects, respectively. The study provides a reference for river nutrients management in arid oases with irrigated agriculture and a scientific and targeted basis to mitigate water pollution and eutrophication of rivers in arid lands.