北方典型水库型水源地水体叶绿素a含量遥感监测研究

遥感监测具有监测区域广、速度快、省时省力等优势,常用于大面积水质监测。其中,叶绿素a浓度是水质监测的一个重要指标。叶绿素a遥感反演的关键是建立遥感数据和叶绿素a含量的定量关系。本文选取石家庄市饮用水源地黄壁庄水库为研究区域,通过进行水体反射光谱现场测量和同步水质采样,得出研究区水体反射光谱特征,并采用丙酮分光光度法测得各水样中叶绿素a含量。在此基础上通过波段比值模型及一阶微分模型分别分析叶绿素a浓度与反射率之间的线性相关性。结果表明,黄壁庄水库各采样点水体叶绿素a浓度均在4.55μg·L-1以下,光谱反射率比值R705nm/R680nm和696 nm反射率的一阶微分值均与叶绿素a浓度有较显著的线性相关性(r2分别为0.736 6和0.875 5)。本文所得结论与方法可为未来对北方水库型水源地水体进行大面积遥感监测叶绿素a含量提供理论参考。     

激光诱导荧光水体叶绿素a浓度监测仪器研制

叶绿素a浓度是反映水质的重要参数之一，常作为水产养殖、渔场预测、环境保护等领域的重要评价指标。针对现有叶绿素a监测仪器实时性差、自动化程度低、环境干扰严重等问题，该研究以实时监测叶绿素a浓度为主要目标，提出了环境光校正算法和数据处理算法，设计了一种基于激光诱导荧光技术的水体叶绿素a浓度监测装置。该装置可向水体发射波长440 nm的激光，诱导激发叶绿素a产生中心波长680 nm的荧光，利用可见光传感器获取叶绿素a受激产生的680 nm荧光强度，即可实现对叶绿素a浓度的监测。以绿藻门小球藻和蓝藻门铜绿微囊藻为对象，利用680 nm荧光值和叶绿素a浓度的线性关系建立拟合模型，模型决定系数R²为0.99；基于统计分析法设计的数据处理可有效剔除误差数据，提高测量结果的稳定性；利用环境光校正算法可有效降低环境光对测量结果的干扰。在仪器稳定性试验中，通过测量激光模块的供电电流和光谱模块的440 nm通道光强值，4 min后两者的标准差系数均为0.001，皮尔逊相关系数为0.84，表明可通过控制电流强度控制激光输出。在性能验证试验中，2组不同光照强度和6组不同藻种的待测水样的平均测量误差在5.48%以内。研究结果可为水质监测领域提供设备支持。     

太湖南泉水域水体及水华蓝藻中常量元素Ca Na Mg K和Al的特征和变化

近年来以微囊藻为优势种的太湖蓝藻水华持续暴发,已经成为严重的环境问题之一。本文通过现场采样及室内分析,研究了2009年太湖南泉水域蓝藻发生过程中藻类群落组成的变化,并同步研究了蓝藻体内及其水环境中常量元素Ca、Na、Mg、K和Al元素浓度的周年变化,分析了三者之间可能的相关性。研究显示,太湖蓝藻发生时间为6—10月,期间水温保持在20℃以上,pH值在水华发生后期可升高至9.8;水体中元素浓度依次为Na〉Ca〉Mg〉K〉Al,而水华蓝藻中元素浓度依次为Ca〉K〉Mg〉Na〉Al;藻体对水体环境中元素的富集系数大小依次为Al〉K〉Mg〉Ca〉Na。研究结果既为选择最佳物理除藻时期来有效降低湖泊中常量元素的负荷量,从而进行水华控制提供了支撑,也为今后将富含元素的水华蓝藻作为＂生物矿源＂加以利用提供了理论依据。     

The Effect of Bioavailable Sedimentary Iron on the Growth of Cyanobacteria in Eutrophic Lakes

The objectives of this study were to investigate whether sedimentary iron can promote the growth of cyanobacteria and determine the effects of different forms of iron on cyanobacterial growth. In this study, we simulated cyanobacterial growth in a eutrophic freshwater lake under Fe-deficient conditions using three systems containing artificial lake water and algae (the WA system); artificial lake water and sediment (WS system); and artificial lake water, sediment and algae (the WSA system). Results demonstrate that Fe from sediments did facilitate cyanobacterial growth. Sequential Fe extractions revealed that the majority of sedimentary iron was in the form of reactive iron (80.63%). Furthermore, cellular iron from cyanobacteria and water-soluble Fe in sediments had a strong and significant negative correlation (??0.792, P?<?0.01), indicating that water-soluble Fe in sediments is the most important form of iron for influencing cyanobacterial growth. Further studies on water-soluble Fe mobility showed that up to 47.5% of the released water-soluble Fe could be absorbed into cyanobacterial cells, thereby indicating that water-soluble Fe is significant for cyanobacterial growth and serves as a critical Fe source when lake water iron is limited. In addition, the study found that easily reducible Fe oxide minerals have the largest release potential. These findings provide new insights that could improve management of cyanobacterial blooms.     

利用BP神经网络模型对太湖水体叶绿素a含量的估算

利用太湖2001-2006年常规水质监测资料和气象资料进行主成分分析,确定影响太湖水体叶绿素a 含量的主要因子。在此基础上构建BP神经网络模型,利用模型对太湖湖心区水体叶绿素a含量进行估算,并对模型进行敏感度分析;将所建模型运用于太湖梅梁湾、贡湖湾、竺山湾以及东太湖4个湖区叶绿素a含量的估算,以验证其适用性。结果表明：基于主成分分析的BP神经网络模型估算的湖心区叶绿素a含量与实测值的拟合度良好,对已建立的BP神经网络预测模型进行敏感度分析表明,气温和溶解氧与浮游植物叶绿素a含量密切相关;该模型对太湖其它4个湖区水体叶绿素a含量的估算结果与实测值拟合度良好, 表明其适用性也较好,因此,可以运用于对太湖水体叶绿素a含量的估算及预测。     

Hyperspectral Remote Sensing of Total Phosphorus (TP) in Three Central Indiana Water Supply Reservoirs

The connection between nutrient input and algal blooms for inland water productivity is well known but not the spatial pattern of water nutrient loading and algae concentration. Remote sensing provides an effective tool to monitor nutrient abundances via the association with algae concentration. Twenty-one field campaigns have been conducted with samples collected under a diverse range of algal bloom conditions for three central Indiana drinking water bodies, e.g., Eagle Creek Reservoir (ECR), Geist Reservoir (GR), and Morse Reservoir (MR) in 2005, 2006, and 2008, which are strongly influenced anthropogenic activities. Total phosphorus (TP) was estimated through hyperspectral remote sensing due to its close association with chlorophyll a (Chl-a), total suspended matter, Secchi disk transparency (SDT), and turbidity. Correlation analysis was performed to determine sensitive spectral variables for TP, Chl-a, and SDT. A hybrid model combining genetic algorithms and partial least square (GA-PLS) was established for remote estimation of TP, Chl-a, and SDT with selected sensitive spectral variables. The result indicates that TP has close association with diagnostic spectral variables with R ² ranging from 0.55 to 0.72. However, GA-PLS has better performance with an average R ² of 0.87 for aggregated dataset. GA-PLS was applied to the airborne imaging data (AISA) to map spatial distribution of TP, Chl-a, and SDT for MR and GR. The eutrophic status was evaluated with Carlson trophic state index using TP, Chl-a, and SDT maps derived from AISA images. Mapping results indicated that most MR belongs to mesotrophic (48.6%) and eutrophic (32.7%), while the situation was more severe for GR with 57.8% belongs to eutrophic class, and more than 40% to hypereutrophic class due to the high turbidity resulting from dredging practices.     

富营养化浅水湖泊蓝藻的种群结构和多样性研究

比较研究不同富营养化浅水湖泊中蓝藻种群结构和多样性,对于认识蓝藻水华的区域分布特征具有重要的生态和环境意义。于2009年7月在富营养化严重的滇池、巢湖和太湖各选取4个采样点,采用16S-23SrRNA基因间隔区（Internaltranscribedspacer,ITS）序列作为生物标记,通过末端限制性片段长度多态性技术（Terminal restriction fragment length polymorphism,T-RFLP）分析蓝藻种群结构和多样性。实验共得到36个不同的末端限制性酶切片段（Terminal restriction fragment,T-RF）,反映3个湖泊中蓝藻种群具有丰富的基因多样性。T-RFLP聚类分析结果表明,在所研究的湖泊中蓝藻种群结构存在空间差异性,其中滇池和巢湖间蓝藻种群结构更为相似。环境因子与蓝藻种群的多元统计分析表明,正磷酸盐（POa-P）和pH与附着细菌样品蓝藻种群结构动态变化具有显著的相关性（P〈0．05）,电导率（Conductivity）和总有机碳（TOC）与浮游细菌样品蓝藻种群结构动态变化具有显著的相关性（P〈0．05）。因此,不同湖泊蓝藻水华的基因型和驱动因子存在差异。     

基于高光谱数据的水体叶绿素a指数反演模型的建立

水体叶绿素a含量是反映水体质量的重要指标之一,利用遥感技术监测其含量具有众多优势。该研究利用2012年7月在广西壮族自治区桂林市漓江流域实地采集的水体高光谱数据和实验室化验分析数据,借鉴陆表植被叶绿素a的遥感反演模型,发展了一种新的水体叶绿素a提取指数(water chlorophyll-a index,WCI)。通过与反射率敏感波段法、波段比值法和半分析方法对比分析发现,新提出的WCI指数使用650、685、696 nm波段,波段稳定,决定系数R2可达0.58,均方根误差最小为0.24,受水体悬浮物影响小,在天津海河区域的验证效果也表明了该模型可以有效地提取水体叶绿素a含量。该方法扩展了水体叶绿素a监测的建模思路,对水体叶绿素a监测建模有一定的指导作用。     

Development of cyanobacterial blooms in Valencian rice fields

Seasonal variation of biomass, photosynthetic pigments and C and N contents of the cyanobacterial blooms developed during the crop cycles of 1998, 1999 and 2000 at two locations in Valencian rice fields (Spain) were studied to find their potential contribution to soil fertility. Blooms of Gloeotrichia sp., Gloeocapsa sp., Microchaete sp., and Nostoc sp. were small and dispersed and appeared only for a few weeks in the experimental fields of the Tancat de Malta location during the crop season of 1998. The biomass of these blooms ranged from 4.4 to 12.8 mg dry wt cm^–2. A bloom of Anabaena sp. was found in the same location during most of the crop cycles of 1998 and 1999, covering up to 80% of the experimental fields. The biomass of this bloom ranged from 2.7 to 11.4 mg dry wt cm^–2. In the moment of its maximal extension (July 1998) we estimate a total biomass of 2,100 kg dry wt for this bloom, equivalent to 420 kg dry wt ha^–1. In the Sueca location a bloom of Microchaete sp. was observed in August 1999 and during most of the crop cycle of 2000, when it covered a wide area (60–70%) of the experimental field. The biomass of this bloom ranged from 0.9 to 5.6 mg dry wt cm^–2. Carbon contents of the blooms ranged from 150 to 310 g mg^–1 dry wt and N contents ranged from 11 to 30.2 g mg^–1 dry wt. Taking into account their N content and their extension we estimate that the incorporation of cyanobacterial blooms could add 4–12 kg N ha^–1 to soil. From an agronomical point of view this means that, in spite of their impressive aspect, cyanobacterial blooms can only fulfill a small proportion of the N requirements of rice plants.     

An Application of Landsat-5TM Image Data for Water Quality Mapping in Lake Beysehir, Turkey

The main goal of this study was to investigate spatial patterns in water quality in Lake Beysehir, which is the largest freshwater reservoir in Turkey, by using Landsat-5TM (Thematic Mapper) data and ground surveys. Suspended sediment (SS), turbidity, Secchi disk depth (SDD), and chlorophyll-a (chl-a) data were collected from 40 sampling stations in August, 2006. Spatial patterns in these parameters were estimated using bivariate and multiple regression (MR) techniques based on Landsat-5TM multispectral data and water quality sampling data. Single TM bands, band ratios, and combinations of TM bands were estimated and correlated with the measured water quality parameters. The best regression models showed that the measured and estimated values of water quality parameters were in good agreement (0.60 < R ² < 0.71). TM3 provided a significant relationship (R ²?=?0.67, p?<?0.0001) with SS concentration. MR between chl-a and various combinations of TM bands showed that TM1, TM2, and TM4 are strongly correlated with measured chl-a concentrations (R ²?=?0.60, p?<?0.0001). MR of turbidity showed that TM1, TM2, and TM3 explain 60% (p?<?0.0001) of the variance in turbidity. MR of SDD showed a strong relationship with measured SDD, with R ²?=?0.71 (p?<?0.0001) for the ratio TM1/TM3 and TM1 band combinations. The spatial distribution maps present apparent spatial variations of selected parameters for the study area covering the largest freshwater lake and drinking water reservoir in Turkey. Interpretation of thematic water quality maps indicated similar spatial distributions for SS, turbidity, and SDD. A large area in the middle portion of the lake showed very low chl-a concentrations as it is far from point and nonpoint sources of incoming nutrients. The trophic state index values were calculated from chl-a and SDD measurements. Lake Beysehir was classified as a mesotrophic or eutrophic lake according to chl-a or SDD parameters, respectively.     

In Situ partitioning and biomagnification of mercury in industrially polluted Husainsagar Lake,Hyderabad, India

The distribution of Hg in water, sediment, plankton and fish along with the in situ biomagnification of the metal in Husainsagar Lake, India were measured. The concentration of Hg in the water was 10 μg L^?1 and exceeded the water quality criteria standards. Sediments of the lake at different sites had Hg concentrations several thousand fold more than that of the overlying water column and showed relationship with the water content of the wet sediment. Sediments in the lake acted as a `sink' for the metal. The concentrations of Hg increased from nanoplankton → phytoplankton -> zooplankton. Fish, an end organism in the food chain, had Hg concentrations higher than that of nanoplankton and lower than those of phytoplankton and zooplankton. This trend did not illustrate the expected pattern of food chain enrichment in the classical sense as noted for chlorinated hydrocarbons (DDT).     

Planktonic algal bloom significantly alters sediment bacterial community structure

Purpose

We determined the relationship of a planktonic algal bloom with spatial and temporal changes in sediment bacterial communities in a eutrophic urban river in the Taihu Basin, China.

Materials and methods

Surface sediments from different locations on the river, including with and without an algal bloom, were collected monthly for 1 year. Sediment bacterial communities were assessed by sequencing 16S rRNA gene amplicons using an Illumina MiSeq.

Results and discussion

There were distinct spatial and temporal changes in sediment bacterial community structure. Significant decrease in the Shannon diversity corresponded with the peak chlorophyll a (Chl a) concentration. Proteobacteria, Chloroflexi, and Bacteroidetes were the dominant phyla throughout the year; however, their relative abundances changed seasonally. At the time of peak Chl a concentrations, Proteobacteria and Bacteroidetes comprised a greater proportion of the bacterial community. Bacterial community structure also varied spatially at the operational taxonomic unit (OTU) level according to canonical correspondence analysis (CCA), especially in locations with an algal bloom compared with those without an algal bloom. There were positive correlations between multiple bacterial genera and Chl a content, suggesting the potential for facilitative relationships between phytoplankton and bacteria. Based on the CCA, water temperature, NH₄ ⁺-N, TN, NO₃ ^?-N, and TP in the sediment and Chl a contents in the water column were significantly correlated with sediment bacterial community structure (P < 0.05).

Conclusions

Planktonic algal blooms may influence sediment bacterial community structure at all taxonomic levels in urban rivers. This work emphasizes the need for more comprehensive studies regarding the impact of planktonic algal blooms on sediment bacterial communities.

     

藻华暴发期太湖不同湖区水样对斑马鱼胚胎发育及仔鱼生长毒性研究

为了评价太湖蓝藻暴发对生态系统的危害程度,于蓝藻水华暴发的2009年7月在太湖梅梁湾（M1和M2）、湖心区（H）和胥口湾（X）4个点采集水样,将斑马鱼（Danio rerio）胚胎直接暴露于水样中,研究太湖不同湖区水质对斑马鱼胚胎发育和仔鱼生长的影响。结果表明,暴露于梅梁湾M2点的斑马鱼胚胎畸形率为7%,显著高于空白组（K）（P〈0.01）,湖心H和梅梁湾M1点为5%左右,也高于空白组（P〈0.01）,而胥口湾X点和空白组无显著性差别（P〉0.05）。蓝藻暴发水体对斑马鱼胚胎产生致畸作用,表现为脊椎弯曲、尾部弯曲和心包水肿等。同时,不同湖区水样对仔鱼生存活力指数（SAI）的影响表明,梅梁湾M2点仔鱼活力最弱,胥口湾X点仔鱼活力最强。试验期间梅梁湾M2点蓝藻密度最高,胥口湾最低,太湖蓝藻密度的分布与斑马鱼胚胎发育的畸形频率具高度的一致性。因此,藻华暴发水体会对脊椎动物胚胎发育和仔鱼生长产生不利影响,危害水生态系统的安全。     

贵州高原红枫湖水库叶绿素a浓度的时空分布及其与环境因子关系

采用2010年对红枫湖水库叶绿素a和理化因子的逐月监测数据,分析了叶绿素a浓度的分布、动态及其与环境因子的关系。结果表明：红枫湖水库叶绿素a浓度具有明显的时空分布特征,在时间上,叶绿素a浓度排序为夏季〉秋季〉冬季〉春季,且在7月份最高,原因是降雨携带充足的磷进入红枫湖,使浮游植物迅速增殖;在空间上,叶绿素a浓度沿入湖河流至大坝出水逐渐降低,总体上南湖高于北湖,这与红枫湖水库磷污染源的空间分布以及浮游植物生长为磷限制有关,南湖因上游有工业点源和城市生活废水的输入而使磷浓度高于北湖。水体叶绿素a与透明度在春季、夏季和冬季呈负相关,与水温在夏季呈正相关,与总磷在春季和秋季呈正相关,而与总氮相关性较复杂,与氨氮无相关性。叶绿素a浓度水平指示红枫湖水库水体处于富营养化状态。     

Urea in the Tributaries of the Chesapeake and Coastal Bays of Maryland

Concentrations of dissolved urea were monitored in several Chesapeake Bay tributaries from 1998 to 2002. Urea is a commonly used agricultural fertilizer and is also a breakdown product of poultry manure, which is used as an additional source of fertilizer throughout the watershed. Two trends were apparent. First, in several of the tributaries, seasonal peaks in ambient urea concentration coincided with the periods of the year (early spring and mid summer) when agricultural applications are most common. Second, highest annual mean concentrations (up to 2.6 μg atom N L^?1), as well as highest individual measurements (up to 24 μg atom N L^?1), were found for those tributaries with the most intensive agricultural and poultry operations. Peak urea concentrations were significantly higher than those which could be attained from in situ sources and regeneration. These elevated concentrations are of concern because this form of nitrogen has been shown to be a preferred form of nitrogenous nutrient for many phytoplankton, including some dinoflagellates which form harmful algal blooms. These results demonstrate that urea from land based sources can contribute to anthropogenic eutrophication.     

高寒草地群落物种多样性与土壤环境因子的关系

对新疆巴音布鲁克高寒草地植物群落4个物种多样性指数和14个土壤指标进行回归分析,结果表明,全盐,HCO3-,Ca2+,速效N,pH值,土壤含水量与物种多样性有显著相关关系(P<0.05).全盐,HC3-,Ca2+,速效N和土壤含水量与丰富度指数的最好拟合为二项式,pH值与丰富度指数的最好拟合为直线回归,全盐,Ca2+.和pH值与Shannon-Wiener指数的最好拟合为二项式,土壤含水量与Shannon-Wiener指数的最好拟合为直线回归,这表明物种多样性与土壤环境因子的关系并不是简单的正相关或负相关关系.     

Impact of Environmental Conditions on the Phytoplankton Structure in Mediterranean Sea Lagoon, Lake Burullus, Egypt

Lake Burullus is one of the northern Delta lakes, connected with the Mediterranean Sea through El-Boughaz opening. The main objective of the present work is to study the physical and chemical properties of the lake and its relation with the phytoplankton communities, chlorophyll-a as well as diversity and Eutrophication index. The studies revealed that, the pH values lie in alkaline side, with wide range of fluctuation (7.15–8.5), the total suspended matter was relatively high (18.2–149 mg l^?1), high values of dissolved oxygen (5.17–11.29 ml l^?1) and high concentration of nutrient salts, (nitrate ranged between (3.4–44.7 μmole l^?1), nitrite (0.42–15.95 μmole l^?1), ammonia (1.46–50.60 μmole l^?1), phosphate (0.63–14.83 μmole l^?1), and silicate (6.54 to 119.3 μmole l^?1). Chlorophyll-a concentration showed a wide range of fluctuation (13.8–127.4 μg/l) and the highest value were recorded at the western region of the lake. Phytoplankton counts showed a pronounced increase (1.9 × 10⁶ units l^?1) as compared with previous studies, a total of 170 species were identified represented mainly by Bacillariophyceae (44.8% to the total community) comprising 68 species, Chlorophyceae (39.99%) 54 species, followed by Cyanobacteria (9.52%) 26 species, Euglenophyceae (5.63%) 15 species. Phytoplankton standing crop attained the highest counts at the western sector particularly at station X during spring and summer (6.7×106 and 4.1×10⁶ units l^?1 respectively) due to the dominance of Bacillariohycean species Stephanodiscus phytoplankton diversity was high and showed widely range (1.47–3.66 nats). A series of stepwise regression equations describing the dependence of phytoplankton standing crop, its main groups and diversity index on the changes of the most biotic prevailing conditions were given and discussed. In general, the results showed that the Lake is considered as a eutrophic area. This phenomenon observed in the body of the lake water that receives large influxes of nutrients, which reflects high phytoplankton counts, diversity index and Chlorophyll-a     

水稻微分光谱和植被指数的作用探讨

通过两年两个不同品种水稻在不同供氮水平下的田间试验，测定了水稻冠层在生长过程中不同时期的高光谱反射率及对应的叶绿素、类胡萝卜素含量，分析了微分光谱对消除水稻冠层光谱的背景影响和植被指数在农学参数测定中的作用。结果表明：由微分光谱所得的红边位置、红边斜率与盖度和供氮水平之间有一定的相关性，与叶面积指数、地上生物量及冠层叶绿素、类胡萝卜素含量有显著相关；水稻多光谱植被指数RVI、NDVI与叶面积指数LAI及其地上部生物量之间有极显著相关性；高光谱植被指数及其变量与植被盖度、供氮水平、叶面积指数、地上生物量及叶绿素、类胡萝卜素含量之间存在相关性。这表明，用微分光谱技术与植被指数方法来监测水稻的色素含量和长势应该是可行的。     

PCR扩增法检测江苏省5大淡水湖泊产毒微囊藻的空间分布

微囊藻毒素（Microcystin,MC）的产生受微囊藻毒素合成酶基因簇（microcystin biosynthesis gene,mcy）调控,常用PCR扩增mcy基因检测产毒微囊藻。采集江苏省5大淡水湖泊——太湖、滆湖、高宝-邵伯湖、洪泽湖和骆马湖的水样,测定水体营养盐浓度和叶绿素a浓度,根据叶绿素a浓度计算5个湖泊的富营养化指数（Trophic state index,TSI）,同时应用单一和多重PCR扩增mcy基因。结果表明,太湖和滆湖处于富营养和超富营养化水平,洪泽湖和骆马湖处于中营养和富营养化水平,高宝-邵伯湖处于寡营养水平。太湖、滆湖、洪泽湖和骆马湖的所有水样均检出mcy基因,4个湖泊水体受到微囊藻毒素的潜在威胁,高宝-邵伯湖没有检测出mcy基因的存在,尚未受微囊藻毒素的污染。     

Numerical modeling of nutrient transport to assess the agricultural impact on the trophic state of reservoirs

A trophic state increase relates to surface water bodies nutrient enrichment, due to the chemical products used such as fertilizers in agriculture and residues from cattle raising activities. This research consists of nutrient transport numerical modeling to analyze the Betancí reservoir trophic state in Colombia; Water samples were collected to analyze total nitrogen, total phosphorus, ammonia, nitrates, nitrites, phosphates, chlorophyll-a, dissolved oxygen, BOD, COD, suspended solids, and water transparency. The water quality model MOHID Studio was implemented and after its calibration, scenarios of increase and decrease of nutrients and inflows to the reservoir through its main tributaries were simulated to reproduce the agricultural activity changes in the basin and the effects that would have on the reservoir's trophic state dynamics; Therefore, the Carlson Trophic Status Index was calculated for each case. The results show that variations in total nitrogen concentration and increased inflows present short-term consequences on the reservoir's trophic state. Increasing the incoming total nitrogen concentrations by 100% causes the reservoir to change from a light eutrophic to a hypereutrophic state. The results of this research provide a starting tool to water resources integrated management in reservoirs.