长江中下游部分湖泊沉积物碱性磷酸酶分布及其作用研究

沉积物磷负荷在湖泊富营养化的发生与恢复过程中具有关键作用,其释放受物理、化学与生物机制调节,而碱性磷酸酶催化有机磷的矿化,故当为促进沉积物磷循环的重要因素。本文讨论了长江中下游部分湖泊沉积物碱性磷酸酶分布及其在磷释放过程中的作用。五里湖疏浚与未疏浚区以及太湖、巢湖、龙感湖、东湖、月湖、龙阳湖、莲花湖等不同湖泊的不同区域表层沉积物碱性磷酸酶活性(APA)明显不同,这种空间异质性与湖泊富营养化程度相联系。此外,APA随沉积物的深度递减,或在中间与较深层次出现峰值,且具明显的季节性。上述事实以及APA对抑制剂的不同响应方式暗示酶存在形态的多样性(同工酶)。苯丙氨酸(Phe)明显提高月湖与五里湖沉积物APA,沉积物与Phe相互作用并静置一天之后,生物可利用性磷(SRP)的释放量明显增加。再者,Phe可抑制月湖沉积物APA,沉积物与Phe相互作用并静置一天之后,SRP释放量无明显变化,溶解有机磷(DOP)的释放量则明显增加。因此,释放的SRP部分来自某些活跃的有机磷的酶促水解,沉积物碱性磷酸酶在内源磷的释放以及富营养化过程中具有重要作用。     

鱼类饲养对地下水中溶解态磷酸酶的影响

比较了鱼类养殖前后 ,地下水中正磷酸盐 (o P)浓度、碱性磷酸酶活性 (APA)在不同大小颗粒之间的分布、溶解态APA对pH、温度、CuSO4、ZnSO4、EDTA 2Na与表面活性剂 (CTAB与TritonX 10 0 )的应答方式及其动力学特征。养鱼之后 ,玻璃缸水中碱性磷酸酶表现出明显较高的活性 ,且以溶解态为主要存在形式 ,这种效应与鱼类的品种有关 ,溶解态APA的最大反应速度 (Vmax)与米氏常数 (Km)均明显提高 ,最适温度与pH值以及对于Zn2 的应答方式亦发生明显改变 ,颗粒结合态APA甚微或不可监测 ,藻类极少 ,故非溶解态APA的主要贡献者。因此 ,鱼类饲养与溶解态APA之间具有密切联系。养鱼缸底颗粒的蒸馏水提取液与同缸水中的溶解态APA对pH值的应答方式十分接近 ,这一结果暗示养殖产生的固体废物是溶解态APA的来源之一。     

池塘水华与底层磷营养状态的关系

比较了武汉东郊相邻池塘间隙水、上覆水与沉积物中磷形态、水相中与不同大小颗粒相联系的碱性磷酸酶活性 (APA)、溶解态与沉积物APA动力学参数及沉积物有机质的含量。铜绿微囊藻 (Microcystisaeruginosa)出现或繁盛的池塘上覆水中正磷酸盐 (O—P)或间隙水中不同形态磷的浓度较高 ,而底层水相不同形态磷浓度较低的池塘叶绿素的浓度亦低 ,且未见铜绿微囊藻 ,故水华与底层磷营养之间联系紧密。各塘沉积物磷主要以铁结合态形式存在 ,铜绿微囊藻繁盛的池塘沉积物中铁结合态磷含量较低 ,而有机质含量较高 ,底层APA亦表现出较高的底物亲和能力 (较小的Km 值 )和反应速度 (较大的Vmax值 ) ,上述结果说明 ,有机质可能在上覆水、间隙水与沉积物诱导具不同动力学特征的高APA ,同时引发厌氧状态 ,这双重因素均有利于O—P的迅速释放 ,从而促进水华的形成     

三峡水库营养元素的分布及其与藻类生长的关系

为进一步研究三峡水库(尤其是支流)氮和磷等营养元素的时空分布格局及其与藻类生长的关系, 于2012 年8 月、11 月与2013 年1 月、4 月在三峡库区木洞至秭归的5 个断面采样分析了不同形态氮和磷以及叶绿素a(Chl.a)的浓度, 并对藻类进行了胞外碱性磷酸酶的荧光标记(Enzyme Labelled Fluoresce, ELF)。结果表明, 春夏季总磷(TP)与Chl.a 浓度均高, 但二者的正相关性在空间尺度上并未得到充分的体现, 干流的TP浓度显著高于支流(P0.05), 而Chl.a 浓度则反之。巫山断面干流与支流的TP 浓度显著低于其他断面(P0.05),而Chl.a 浓度则无显著差异, 秭归断面显著增高的TP 浓度改变了其沿水流方向下降的趋势。从总体上讲,Chl.a 浓度与TP、溶解性总磷(DTP)、溶解反应性磷(SRP)和硝态氮(NO3--N)浓度均极显著负相关(P0.01), 而与总氮(TN)浓度、TN/TP、NO3--N/SRP 等均极显著正相关(P0.01), 因此, 氮对藻类的生长可能具有一定的促进作用, 而泥沙对磷的吸附与沉降有效降低了其生物可利用性。巫山断面底层显著较高的TP 与颗粒态磷(PP)浓度支持了上述推测, 不同断面不同藻类胞外磷酸酶的标记进一步说明了磷对其生长的关键限制性作用。因此, 在水质管理、富营养化防治和渔业资源管理过程中必须充分考虑不同营养元素的存在状态及其与水文条件的交互作用。       

尖刺拟菱形藻和抑食金球藻碱性磷酸酶生理学特性的比较研究

采用批量培养的方法, 比较研究尖刺拟菱形藻 (Pseudo-nitzschia pungens)和抑食金球藻(Aureococcus anophagefferens)两种海洋微藻碱性磷酸酶(Alkaline phosphatase, AP)的生理学特性, 分析细胞内、外磷含量的变化对AP 表达的调控。结果表明, 尖刺拟菱形藻和抑食金球藻的AP 均为诱导酶, 藻的碱性磷酸酶活性(AP activity, APA)表达受到外部溶解态无机磷酸盐(Dissolved inorganic phosphate, DIP)与细胞内颗粒磷含量(Particulate phosphorus, PP)的共同调控。尖刺拟菱形藻APA 对磷胁迫的灵敏度较高, 在DIP 尚高(1.80 μmol⋅L–1)且PP 充分(222.19 fmol⋅cell–1)的条件下, 尖刺拟菱形藻就开始大量表达APA。而抑食金球藻则是在内外磷源都即将耗尽(DIP 和细胞PP 分别为0.26 μmol⋅L–1 和4.49 fmol⋅cell–1)的条件下才开始大量表达APA。尖刺拟菱形藻的AP 基本是结合在细胞上的, 而抑食金球藻会释放一定量(6.1%-20.9%)的AP 于水体中。抑食金球藻单位体积的最大APA 约为尖刺拟菱形藻的73 倍。尖刺拟菱形藻与抑食金球藻依靠AP 水解溶解态有机磷(Dissolved organic phosphorus, DOP)的能力有差异。     

三种水华蓝藻对不同磷浓度生理响应的比较研究

本实验研究了铜绿微囊藻(Microcystis aeruginosa FACHB469)、水华鱼腥藻(Anabaena flos-aquae FACHB245)和浮游颤藻(Oscillatoria planctonicaFACHB708)对磷浓度变化的生理响应。结果表明,在缺磷条件下,A. flos-aquae对低磷环境的适应能力较强,O. planctonica其次,M. aeruginosa最差;在磷充足条件,微囊藻对磷过量吸收的能力明显高于其他两种蓝藻。三种蓝藻胞外碱性磷酸酶活性(APA)与培养基中磷浓度呈负相关性,其产生碱性磷酸酶(AP)的能力由高至低为:A. flos-aquae>O. planctonica>M. aeruginosa。磷缺陷时A. flos-aquae产生的胞外APA约是M. aeruginosa的10倍,是O. planctonica的5倍。     

有机磷农药对铜绿微囊藻生长及摄磷效应的动力学研究

研究了两种有机磷农药 (甲胺磷、辛硫磷 )在较低浓度时对铜绿微囊藻的生长效应以及微囊藻摄取磷形态 :总溶解磷 (TSP)、溶解反应磷 (SRP)和溶解有机磷 (DOP)的动力学规律。保持各培养基中总磷的浓度 6 95mg/L ,与培养基中不加有机磷农药相比 ,当培养基中甲胺磷与K2 HPO4的浓度比为 1∶11和 1∶9,辛硫磷与K2 HPO4的浓度比为1∶349和 1∶114时 ,微囊藻的生长受到促进 ,并且当甲胺磷与K2 HPO4,的浓度比为 1∶9,辛硫磷与K2 HPO4的浓度比为1∶114时促进效果最好 ,而当培养基中没有K2 HPO4甲胺磷作为惟一磷源时 ,微囊藻的生长受到明显的抑制。微囊藻摄取SRP的速度快于其他磷形态 ,而对DOP的利用较少。微囊藻生长越快 ,对SRP的摄取速率也越快。结果表明低浓度有机磷农药促进微囊藻生长的主要原因是有机磷的加入促进了藻类对SRP的摄取     

东北丘陵区林地转型耕地对土壤编码碱性磷酸酶基因的细菌群落的影响

【目的】通过研究林地转型耕地对土壤编码碱性磷酸酶基因的细菌群落丰度、多样性和结构的影响,为丘陵区耕地长期施肥下农田土壤微生物多样性丧失的影响机制以及未来的退耕还林过程中土壤微生物多样性的提升和土地可持续利用研究提供一些基础数据和技术支撑。【方法】采用实时荧光定量PCR (real-time quantitative PCR,qPCR)和高通量测序技术解析土壤编码碱性磷酸酶基因的细菌群落的丰度、多样性和结构变化,并耦合土壤化学性质分析,明确土壤编码碱性磷酸酶基因的细菌群落丰度和多样性与土壤化学性质的关系以及关键的驱动因子。【结果】林地垦殖为农田后,长期施肥导致土壤酸化,pH从5.58降至4.72,而土壤速效磷则从2.49 mg/kg增至49.3 mg/kg。相应地,耕地土壤编码碱性磷酸酶基因的细菌群落的丰度和Shannon指数均显著低于林地。基于编码碱性磷酸酶的phoD基因(alkaline phosphatase-encoding gene)序列的物种分类表明,丘陵区土壤编码碱性磷酸酶基因的细菌群落的优势门为变形菌门(Proteobacteria)、蓝藻门(Cyanobacteria)、浮霉菌门(Planctomycetes)、放线菌门(Actinobacteria)、厚壁菌门(Firmicutes)和疣微菌门(Verrucomicrobia),其中林地土壤的蓝藻门的相对丰度显著高于耕地。耕地土壤的慢生根瘤菌属(Bradyrhizobium)和芽孢杆菌属(Bacillus)的相对丰度显著高于林地,而中慢生根瘤菌属(Mesorhizobium)、假单胞菌属(Pseudomonas)、Chlorogloea属、Gemmata属、Phormidesmis属和Pseudolabrys属的相对丰度显著低于林地。土壤编码碱性磷酸酶基因的细菌群落结构因林地转型耕地而发生显著改变。phoD基因丰度和Shannon指数与pH显著正相关,而与总磷、速效磷、硝态氮和铵态氮均显著负相关,其中土壤速效磷是这些影响因素中影响最强烈的,长期施用无机磷肥导致含碱性磷酸酶的土壤细菌群落对有机磷分解的能力退化。【结论】林地转型耕地加之长期施肥改变了土壤pH和速效磷,并在其他理化因子的协同驱动下,导致土壤编码碱性磷酸酶基因的细菌群落丰度、多样性和结构的显著变化。     

有机磷农药对滇池微囊藻生长和摄磷效应的影响

采集滇池水体作为铜绿微囊藻培养基,研究了两种有机磷农药(甲胺磷和辛硫磷)对微囊藻生长和摄磷效应的动力学规律。结果表明,在滇池水体中添加较低浓度的甲胺磷(0.8、1.6、3.2mg/L)和辛硫磷(0.02、0.06、0.1mg/L)均能不同程度地促进微囊藻的生长,且在HGZ培养基中抑制微囊藻生长的浓度在滇池水体中却能促进微囊藻的生长。微囊藻的生长取决于细胞内磷的浓度且对磷的吸收利用存在积累性,在微囊藻生长初期,摄取各形态磷的速率较快;随后微囊藻摄取各形态磷的速率较慢。总溶解磷(TSP)和溶解反应磷(SRP)是微囊藻优先摄取的磷形态,在生长过程中微囊藻利用了大量的溶解有机磷(DOP)作为磷源加速生长。这一特点对于微囊藻成为淡水湖泊富营养化发展过程中的一种重要优势种具有极为重要的作用。     

不同固磷方式对巢湖沉积物磷吸附行为的影响

内源磷负荷将严重阻碍富营养化湖泊的恢复,其控制技术的关键在于有效增强沉积物吸附磷的能力,而相关研究相对较少。研究以典型富营养化湖泊(巢湖)严重污染区域的沉积物为实验对象,系统比较了常规固磷方式(施用CaCl2、FeCl3、AlCl3与曝气)对沉积物磷吸附行为和间隙水溶解态可反应磷(SRP)浓度的影响。结果表明:施用不同剂量的CaCl2之后,沉积物磷最大吸附量和吸附能均无显著变化,间隙水SRP浓度和沉积物磷平衡浓度(EPC0)仅有较小幅度的下降;FeCl3和AlCl3的施用可明显增加沉积物磷的最大吸附量和吸附能,同时有效降低间隙水SRP浓度和EPC0值,即沉积物显示更强的从水中吸附磷的能力。当同处低剂量水平时,铁能更有效地降低沉积物EPC0值;反之,曝气对沉积物最大吸附量和吸附能均无显著影响,却明显提高了间隙水SRP浓度和沉积物EPC0值,进而导致更强的磷释放风险。故建议将适量铁的施用作为富营养化湖泊沉积物修复的有效技术,且慎用曝气处理。     

Isolation and Characterization of Organic Phosphorus-Mineralizing Bacteria in Sediment of a Chinese Large Shallow Eutrophic Lake (Lake Taihu)

The organic phosphorus mineralizing bacteria (OPB) play an important role in phosphorus cycling in lake sediment, to which less attention has been paid. Monthly sediment samples in 2009 ending in October, together with the samples from different seasons (May, July, October, and December) in 2008, were collected at 6 sites in a Chinese large shallow eutrophic lake (Lake Taihu). The sediment OPB numbers ranged from 2.2 × 10⁶ to 1.79 × 10⁸ cells g^?1 (dry weight) at different sampling sites and in different seasons, with the average being 3.88 × 10⁷ cells g^?1 (dry weight). Its number was highest at the most polluted site and peaked in spring and summer, which can be explained by the enrichment of organic matter in sediment. Furthermore, there existed a significant positive relationship between the OPB numbers and alkaline phosphatase activities in the sediment. The 6 OPB strains isolated from the sediment were distinct in terms of their colony morphology on the yolk agar, biochemical characteristics and phosphorus release abilities. According to the 16S rDNA sequences, these OPB belong to Bacillus cereus, Stenotrophomonas maltophilia, Stenotrophomonas sp., Bacillus cereus, Xanthomonas sp., Pseudomonas sp. They were distinguished from the OPB species recorded in a Chinese small shallow eutrophic lake whose sediment organic content was remarkably higher. Taken together, organic matter shaped the OPB community not only quantitatively but also qualitatively, which in concert facilitated the enzymatic hydrolysis of organic phosphorus in lake sediment.     

Limitations of using extracellular alkaline phosphatase activities as a general indicator for describing P deficiency of phytoplankton in Chinese shallow lakes

Extracellular phosphatase can be produced by phytoplankton to utilize organic phosphorus under phosphorus (P) deficiency. However, there is a controversy about its use as an indicator of P deficiency in natural phytoplankton community inferred by such an “induction–repression” mechanism. Size-fractionation of alkaline phosphatase activity (APA), soluble reactive phosphorus (SRP) concentration, algal density, and composition were determined in six Chinese shallow lakes ranking in gradient of trophic status, where a positive relationship between SRP concentration and algal density was observed. Enzyme-labeled fluorescence (ELF) method was used to localize phosphatase on cell membrane of algae. The so-called algal APA that associated with coarser particle (>3.0 μm) accounted for the largest part of total APA. Within a lake with lower SRP concentration, the “induction–repression” mechanism held true. Contrastingly, both algal and total APA were positively related to SRP concentration based on the data across all study lakes with statistical significance, which may be explained firstly by algal composition. The lakes with higher SRP concentration were dominated by diatoms and green algae, while they easily produced extracellular phosphatases as evidenced by ELFA labeling. In parallel, the lakes with lower SRP concentration were dominated by cyanobacteria, while it was never ELFA-positive; secondly, ELFA-positive dots or structures suggested that, in lakes with higher trophic status, attached bacteria or heterotrophic microorganisms could substantially contribute to extracellular phosphatases for hydrolyzing organophosphoric compounds but probably utilizing the organic moiety as an organic carbon source. This process simultaneously produces inorganic P, leading to the co-occurrence of high phosphate concentration and APA. So, the contributor of APA are complex, which may produce extracellular phosphatase species-specific or not exclusively for P nutrient and consequently make it difficult to normalize APA with the exact biomass estimators. Therefore, it is not reasonable to use APA, normalized or not, as a general indicator for describing P deficiency of phytoplankton in shallow lakes especially eutrophic ones.     

Stratification of alkaline phosphatase in sediments of two urban lakes and its effect on phosphorus cycle

Phosphorus loadings in sediments play an important role in lake eutrophication and the progress of its recovery. The phosphorus release is controlled by physical, chemical and biological mechanisms. Alkaline phosphatase catalyzes remineralization of organic phosphorus and then it may be an important factor accelerating phosphorus cycling in sediments. In this paper, distributions, properties and function of alkaline phosphatase with depths in sediments of two urban lakes were discussed. Alkaline phosphatase activity (APA) in the sediments of Lake Yuehu decreased with the sediment depth. APA in sediments of Lake Yuehu was, mostly, inhibited by Phe and L-Cys; and inhibiting ability of Phe could be stronger than L-Cys. APA in deeper layer (20–30 cm) of sediments was more sensitive to the inhibitors than other layers, but range of variation in APA was most wide in the subsurface layer (10–20 cm). All the facts implied that alkaline phosphatase occurred in various forms (isoenzymes). APA in the sediments with different depths of Lake Donghu responded Phe differently. Reacted with Phe and incubated for 1 day, the amounts of SRP released by these sediments varied correspondingly. SRP on the overlying water in deeper layers (5–10 cm and 15–20 cm) of Site T1 was higher than that in surface layer (0–5 cm) of the same site, 1 day after incubation. Hence, the SRP release resulted, at least partially, from the hydrolysis of some liable organic phosphorus mediated by APA. Alkaline phosphatase in lake sediments plays an important role in the release of internal phosphorus loadings and eutrophication. A possible explanation for the sensitivity at deeper layers could be another active region of hydrolysis by alkaline phosphatase from organic phosphorus, which added a new dimension in phosphorus cycling mediated by some biochemical mechanisms.     

Seasonal Variations in Chlorophyll a Concentrations in Relation to Potentials of Sediment Phosphate Release by Different Mechanisms in a Large Chinese Shallow Eutrophic Lake (Lake Taihu)

The relationship between chlorophyll a and fractionation of sediment phosphorus, inorganic phosphate-solubilizing bacteria (IPB), and organic phosphate-mineralizing bacteria (OPB) was evaluated in a large Chinese shallow eutrophic lake (Lake Taihu) and its embayment (Wuli Bay). At the three study sites, the increase of chlorophyll a concentrations in April paralleled those of the iron bound phosphate accounting for major portion of sediment inorganic phosphate, and in June significantly higher OPB and IPB numbers (especially OPB) in sediment were main contributors to the peaks of chlorophyll a concentration. Even though IPB peaked from February to June, it should serve as an unimportant P source due to the irrelevancy with chlorophyll a and soluble reactive phosphorus (SRP). By contrast, at the other site in the embayment, the calcium-bound phosphate was predominant and solid, which was difficult to be released, and neither IPB nor OPB were detectable in the sediment, indicating weak potential for phosphorus release from the sediment, which was reflected in the small seasonal variation in SRP concentration in water column. Hence, the extents to which the three general mechanisms behind phosphate release from sediment (desorption of iron bound phosphate, solubilization by IPB and enzymatic hydrolysis by OPB) operated were different depending on seasons and sites in Lake Taihu, they may jointly drive phosphate release and accelerate the eutrophication processes.     

Dredging effects on P status and phytoplankton density and composition during winter and spring in Lake Taihu, China

Phytoplankton density and composition, together with phosphorus (P) concentrations and size-fractionated alkaline phosphatase activity (APA), were investigated in dredged and undredged zones in Lake Taihu from January to April 2004. P concentrations were also determined in the corresponding interstitial water. Enzyme Labeled Fluorescence (ELF) was used for localizing extracellular phosphatase on phytoplankton cell membranes in April. The increase in phytoplankton density was paralleled by a significant increase in soluble reactive P (SRP) concentrations in the water column and interstitial water at all sites from January to April, with chlorophyte gradually becoming dominant. In February, at the undredged site, more algae dominated by chlorophyte occurred in overlying water, rather than in the surface, coinciding with higher SRP concentrations in overlying and interstitial water. Therefore, P status in the bottom is important to phytoplankton development in terms of density and composition. Undredged sites had higher SRP concentrations in interstitial water than dredged sites. Furthermore, Higher APA was observed, accompanied by higher dissolved organic P (DOP) and lower total P at the undredged site in February. Enzymatic hydrolysis of DOP may have been an additional source of P for phytoplankton. In April, Schroederia sp. was ELF labeled in surface water at the dredged site, which showed markedly lower SRP concentration, but not at the undredged site with higher SRP concentration. Thus, the dredging might regulate algal density and composition in water column by reducing P bioavailability.     

Comparison of phosphorus deficiency indices during a spring phytoplankton bloom in a eutrophic reservoir

1. Phosphorus limitation was studied along the eutrophic, canyon-type ?ímov reservoir (Czech Republic) during a spring phytoplankton bloom. Concentration of soluble reactive phosphorus (SRP), C:P molar ratio in seston, extracellular alkaline phosphatase activity (APA), and P limitation (bioassay) were used as indices for phosphorus deficiency in the phytoplankton. 2. SRP, C:P, APA, and P limitation indicated a moderate P deficiency in the downstream, but not upper, part of the reservoir. 3. Significant correlations between these parameters were found in the downstream part. Chlorophyll a concentration correlated with APA and P limitation in the upper part. 4. APA was significantly enhanced in the phosphorus-deficient phytoplankton. However, APA was apparently not related to total biomass or species composition of the phytoplankton. 5. Generally, APA was closely correlated with pH in the reservoir. However, extracellular alkaline phosphatases, with a pH optimum above 9.0, were induced and active only during the phytoplankton bloom, whereas low background activity of extracellular phosphatases was found at low chlorophyll a concentrations (winter, clear-water phase).     

不同形态的磷源对球形棕囊藻生长及碱性磷酸酶的影响

研究了不同浓度和形态的无机磷(DIP)和有机磷(DOP)对球形棕囊藻细胞生长、培养基中磷浓度的变化以及碱性磷酸酶活性的变化的影响。在缺磷和大分子DOP(卵磷脂)作为磷源的情况下,碱性磷酸酶活性迅速提高,在第9天达到了最大值,其活性分别为19.11U和18.28U,显然在缺磷胁迫下和大分子DOP的利用过程中碱性磷酸酶起着重要作用。DIP(磷酸二氢钾)和小分子DOP(β-甘油磷酸钠)都容易直接被球形棕囊藻吸收利用,碱性磷酸酶活性变化不明显。     

The phosphorus fractions and alkaline phosphatase activities in sludge

The alkaline phosphatase activities (APA) and phosphorus fractions in activated sludge during wastewater treatment were studied. Our results showed that the phosphorus concentration and fractions in activated sludge were highly correlated with the characteristics of influents. Inorganic phosphorus (IP) and non-apatite inorganic phosphorus (NAIP) were the main phosphorus fractions of sludge. A larger phosphorus concentration was found in activated sludge due to the more readily mobilizable and bio-available forms. The APA in sludge was directly correlated with mixed liquor suspended solids (MLSS) in activated sludge. The APA in the sludge is implicated the depletion of organic phosphorus forms in sludge, whilst also implying its less inhibition of inorganic phosphorus in sludge. The APA and phosphorus fractions in different sludge samples from the same wastewater treatment plant were quite stable. This stability shows their tight interactions in sludge.     

Mineralization of dissolved organic phosphorus from a shallow eutrophic lake

Release of soluble reactive phosphorus (SRP) from dissolved organic phosphorus (DOP), concentrated by reverse osmosis of water samples from Lough Neagh Northern Ireland, was measured in the presence of enzymes and cultures of lake water bacteria in a basal liquid medium adjusted to the pH of lake water (7.6). No hydrolysis of unfractionated DOP was observed in the presence of alkaline phosphatase but a combination of alkaline phosphatase and phosphodiesterase mineralized 14% of DOP in a 30 day incubation period at 15 °C. A similar amount of mineralization was attained by phytase. Phytase induced the same degree of mineralization in a range of DOP fractions varying from MW > 100 000 to c. 500. A mixed culture of lake water bacteria mineralized 12% of unfractionated DOP. Single cultures of lake water bacteria displayed low mineralizing activity (mean of 49 cultures = 5% DOP hydrolysed). Results indicate that DOP from Lough Neagh in the above molecular weight range is predominantly recalcitrant to bacterial mineralization under natural lake conditions.