鱼类饲养对地下水中溶解态磷酸酶的影响
EFFECTS OF FISH CULTURE ON DISSOLVED PHOSPHATASE IN GROUNDWATER
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摘要: 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Triton X-100)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.Abstract: Extracellular phosphatase plays a crucial role in P cycling in aquatic ecosystem. Little attention has been paid on the occurrence of extraccllular phosphatase in groundwater. Furthermore, effects of aquaculture on extracellular phosphatase, especially dissolved alkaline phosphalase,were inadequately studied. The objectivc of this study was to investigate the influences of fish culture on dissolved alkaline phosphatase,including its origin. activity and kinetics,in freshwater. Water was taken from a well,as a control,and 10 different pools rearing a variety of fishes with the same well water in November and December 2001,as well as May 2003,for 5 times. Variables involved in pcycling,such as orthophosphate(o-P)concentrations,size fractionation of alkaline phosphatase. responses of dissolved alkaline phosphatase activity (DAPA)to pH,temperature,CuSO4,ZnSO4,EDTA 2Na. as well as surfactants(CTAB and Triton X-100)and its kinetics,were deternlined. DAPA in groundwater. with and without fish rearing was inhibited by Cu2+ 、CTAB with different concentrations and EDTA-2Na at higher concentration (2mmol/L). At the same time,it was enhanced by Triton X-100 with different concentrations. The sensitive responscs to the spec ificinhibitors provided further evidences that alkaline phosphatase occurred in groundwater. In the water of glass jars with fish culturing,DAPA increased significantly,and the extent of increase was depended on fish species cultured. In details,groundwater with Trichogoster trichopterus and Botia licontei rearing showed significantly higher DAPA than controls. Besides. DAPA was undctectable in the groundwater with Mastcembetus erythrotaenia,Aymphysodon acquifasciata,Monodactylus argentens, Apteronotus aslifrons, Acipenser sinensis Grtey.Cyprinus carpio Linnaeus rearing,but detectable in the water with Gichasoma synspolum,Botia licontei,Mylipharyngodon piceus Richardon,Ctenopharyngodon idellus Cuvier et Valenciennes,Hypophthalmichthys molitrix,Aristichthys nobilis rearin and reshowed the highest value with Trichogoster trichoptern rearing,DAPA was characterized in control and that rearing Trichogoster trichopterus and Botia licontei (culture medium).It showed the highest level at 35℃in control,while it peaked at both 35℃ and 55℃in the culture medium. Its optimum pH was pH 7.0 in control,but was pH 5.8 and pH 8.9 in the culture medium.Moreover,it was enhanced by Zn2+ at higher concentration(0.5mmol/L)in control,but was inhibited significantly by Zn2+ at different concentrations in culture medium.Kinetics of dissolved alkaline phosphatase conformed to the Michaelis Menten model. The significantly higher V max and K m values were recorded in the colture medium compared to those in control.In short,DAPA in culture medium exhibited distinct models responding to temperature,pH ZnSO4 as well as higher V max and K m values. In addition,alkaline phosphatase activity associated with particles(>3.0μm)was very low, or undetectable,coupled with scarcely observed algae cells,indicating that phytoplankton is unlikely to be the main producer of DAPA. Hence the occurrence of DAPA was causatively linked to fish culture. Extracts from particles collected in the bottom of jars with fish culture, by distilled water, exhibited DAPA. It showed a similar manner responding to pH values with that of water in the same jars, implying that the solid wastes resulted from fish culture is one of the sources providing dissolved alkaline phosphatase.
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Keywords:
- Groundwater /
- Fish culture /
- Dissolved phosphatase
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[1] Jamet D,Boge G.Characterisation of marine zooplankton alkaline phosphatase activity in relation to water quality[J].Hydrobiologia,1998,373-374:311-316[2] Wynne D.The role of phosphatase in the metabolism of peridinium cinctum.from Lake Kinneret[J].Hydrobiologia,198l,83:93-99[3] Song C L,Cao X Y,Liu B Q,et al.A relationship between algae blooms and benthic phospharus status in ponds[J].Acta.Hydrobiologica.Sinica.,2004,28(1):7-12.[宋春雷,曹秀云,刘兵钦,等.池塘水华与底层磷营养状态的关系.水生生物学报,2004,28(1):7-12][4] Sinsabaugh,Foreman.Activity profiles of bacterioplankton in a eutrophic river[J].Freshwater Biology,2001.46(9):1239-1249[5] Baneras L,Rodriguez-Gonzalez J,Garcia-Gil L J.Contribution of photosynthetic sulfur bacterra to the alkaline phosphatase activity in anoxic aquatic ecosystems[J].Aquatic Microbial Ecology,1999,18(1):15-22[6] Zhou Y Y Dodds W K.Kinetics of size-fractionated and dissolved alkaline phosphatase in a farm pond[J].Archiv fur Hydrobiologie.Stuttgart,1995,134(1):93-102[7] Zhou Y Y, Li J Q,Fu Y Q.Effects of submerged macrophytes on kinetics of alkaline phosphatase in Lake Donghu.I.Unfiltered water and sediments[J].Water Research,2000,34(15):3737-3742[8] Zhou Y Y,Li J Q,Zhang M.Relationship between the kinetics of alkaline phosphatase and aquatic plants in wetlands[J].Journal of Lake Sciences,2002,14(2):138-143.[周易勇,李建秋,张敏.湿地中碱性磷酸酶的动力学特征与水生植物的关系.湖泊科学,2002,14(2):138-143][9] Zhou Yiyong,Li Jianqiu,Song Chunlei,et al.Seasonality in activity and heterogeneity in composition of free alkaline phosphatase in interstitial water of lake Donghu[J].Acta.Hydrobiologica.Sinica.,2003,27(1):100-102.[周易勇,李建秋,宋春雷,等.东湖间隙水自由碱性磷酸酶活性的季节性与组成的非均一性.水生生物学报,2003,27(1):100-102][10] Currie D J,Bentzen E,Kalff J.Does algal-bacterial phosphorus partitioning vary among lakes? A comparative study of orthophosphate uptake and alkaline phosphatase activity in freshwater[J].Can.J.Fish Aquat Sci,1986,43:311-318[11] Jansson M.Phosphatases in Lake water:Characterization of Enzymes from phytoplankton and Zooplankton by Gel Filtralion[J].Science,1976,194:320-321[12] Healey F P, Hendzel LL.Fluoromeric measurement of alkaline phosphatase activity in algac[J].Freshwater Biol.1979,9:429-439[13] Brown S E,Goulder R.Extraccllular-enzyme activity in trout-farm effuents and a recipient river[J].Aquaculture Research,1996,27(12):895-901[14] Carr O J,Goulder R.Fish-farm effluents in rivers-1.Effects on bacterial populations and alkaline phosphatase activity[J].Water Research 1990,24(5):63l-638[15] Mauraa G, Eville G.Alkaline phosphatase activity and cellular phosphorus as an index of the phosphorus status of phytoplankton in Minnesota lakes[J].Freshwaer Biology,1985,15:227-233[16] Murphy J,Riley P.A modified single solution method for the determination of phosphate in natural waters.[J].Anal Chim Acta,1962,27:31-36[17] Huang Q Y,Haruo Shindo.Comparison of the lnfluence of Cu,Zn,and Cd on the Activity and Kinetics of Free and Immobilized Acid Phosphatase[J].Soil Sci.Plant Nutr.,200l,47(4):767-772[18] Kunito T,Saeki K,Goto S, et al.Copper and zinc fractions affecting microorganisms in long-term sludge-amended soils[J].Bioresour.Technol,200l,79(2):135-146[19] Lalitha J,Mulimani V H.Stabitlity and activity of potato acid phosphatase in aqueous surfactant media [J].Biochemistry and molecular biology international, 1997,41(4):797-803[20] Yang D, Wang J Y,Peng X J,et al.Kinetics of inactivation of Ulva pertusa Kjellm alkaline phosphatase by ethylenediaminetetraacetic acid disodium[J].Journal of Enzyme Inhibition,2001,16(4):331-319[21] Janna B B,Das S K.Bioturbation induced changes of fertilizer value of phosphate rock in retation to alkaline phosphatase activtity[J].Aquaculture,1992,103(3-4):321-330[22] Newman S.Sediment resuspension effects on alkaline phosphatase activity[J].Hydrobiologia,1992,245:75-86[23] Laura S,Paul I B.Effect of polyphenolic compounds on alkaline phosphatase activity:Its implication for phosphorus regeneralion in Australian freshwaters[J].Arch.Hydrobiol.,1991,123(1):1-19[24] Buainain L B,Kaowaki M K,Maria de Lourdes Polizeli: et al.Characterization of a conidial alkaline phosphatase from the thernophilic fungus Humicola grisea var.thermoidea [J].Basic Microbiol.,1998,38(2):85-94[25] Zhou Y Y, Li J Q,Fu Y Q,et al.Kinetics of alkaline phosphatase in lake sediment associated with cage culture of Oreochromis nilotucus [J].Apuaculture, 2001,203:23-32[26] János Oláh,Erzsébet O.Tth.The function of alkaline phosphatase enzyme in the phosphorus cycle of fertized fishponds [J].Aquacultura Hungarica(Szarvas),1978,1:15-23[27] Barik S K,Purushothaman C S,Mohanty A N.Phosphatase activity with refercnce to bacteria and phosphorus in tropical freshwater aquaculture pond systems[J].Aquaculture Research,2001,32(10):819-832[28] Scholz O,Marxsen J.Sediment phosphatases of the Breitenbach (Germany), a first-order central European stream [J].Arch.Hudrobiol.,1996,135(4):433-450 Jamet D,Boge G.Characterisation of marine zooplankton alkaline phosphatase activity in relation to water quality[J].Hydrobiologia,1998,373-374:311-316[2] Wynne D.The role of phosphatase in the metabolism of peridinium cinctum.from Lake Kinneret[J].Hydrobiologia,198l,83:93-99[3] Song C L,Cao X Y,Liu B Q,et al.A relationship between algae blooms and benthic phospharus status in ponds[J].Acta.Hydrobiologica.Sinica.,2004,28(1):7-12.[宋春雷,曹秀云,刘兵钦,等.池塘水华与底层磷营养状态的关系.水生生物学报,2004,28(1):7-12][4] Sinsabaugh,Foreman.Activity profiles of bacterioplankton in a eutrophic river[J].Freshwater Biology,2001.46(9):1239-1249[5] Baneras L,Rodriguez-Gonzalez J,Garcia-Gil L J.Contribution of photosynthetic sulfur bacterra to the alkaline phosphatase activity in anoxic aquatic ecosystems[J].Aquatic Microbial Ecology,1999,18(1):15-22[6] Zhou Y Y Dodds W K.Kinetics of size-fractionated and dissolved alkaline phosphatase in a farm pond[J].Archiv fur Hydrobiologie.Stuttgart,1995,134(1):93-102[7] Zhou Y Y, Li J Q,Fu Y Q.Effects of submerged macrophytes on kinetics of alkaline phosphatase in Lake Donghu.I.Unfiltered water and sediments[J].Water Research,2000,34(15):3737-3742[8] Zhou Y Y,Li J Q,Zhang M.Relationship between the kinetics of alkaline phosphatase and aquatic plants in wetlands[J].Journal of Lake Sciences,2002,14(2):138-143.[周易勇,李建秋,张敏.湿地中碱性磷酸酶的动力学特征与水生植物的关系.湖泊科学,2002,14(2):138-143][9] Zhou Yiyong,Li Jianqiu,Song Chunlei,et al.Seasonality in activity and heterogeneity in composition of free alkaline phosphatase in interstitial water of lake Donghu[J].Acta.Hydrobiologica.Sinica.,2003,27(1):100-102.[周易勇,李建秋,宋春雷,等.东湖间隙水自由碱性磷酸酶活性的季节性与组成的非均一性.水生生物学报,2003,27(1):100-102][10] Currie D J,Bentzen E,Kalff J.Does algal-bacterial phosphorus partitioning vary among lakes? A comparative study of orthophosphate uptake and alkaline phosphatase activity in freshwater[J].Can.J.Fish Aquat Sci,1986,43:311-318[11] Jansson M.Phosphatases in Lake water:Characterization of Enzymes from phytoplankton and Zooplankton by Gel Filtralion[J].Science,1976,194:320-321[12] Healey F P, Hendzel LL.Fluoromeric measurement of alkaline phosphatase activity in algac[J].Freshwater Biol.1979,9:429-439[13] Brown S E,Goulder R.Extraccllular-enzyme activity in trout-farm effuents and a recipient river[J].Aquaculture Research,1996,27(12):895-901[14] Carr O J,Goulder R.Fish-farm effluents in rivers-1.Effects on bacterial populations and alkaline phosphatase activity[J].Water Research 1990,24(5):63l-638[15] Mauraa G, Eville G.Alkaline phosphatase activity and cellular phosphorus as an index of the phosphorus status of phytoplankton in Minnesota lakes[J].Freshwaer Biology,1985,15:227-233[16] Murphy J,Riley P.A modified single solution method for the determination of phosphate in natural waters.[J].Anal Chim Acta,1962,27:31-36[17] Huang Q Y,Haruo Shindo.Comparison of the lnfluence of Cu,Zn,and Cd on the Activity and Kinetics of Free and Immobilized Acid Phosphatase[J].Soil Sci.Plant Nutr.,200l,47(4):767-772[18] Kunito T,Saeki K,Goto S, et al.Copper and zinc fractions affecting microorganisms in long-term sludge-amended soils[J].Bioresour.Technol,200l,79(2):135-146[19] Lalitha J,Mulimani V H.Stabitlity and activity of potato acid phosphatase in aqueous surfactant media [J].Biochemistry and molecular biology international, 1997,41(4):797-803[20] Yang D, Wang J Y,Peng X J,et al.Kinetics of inactivation of Ulva pertusa Kjellm alkaline phosphatase by ethylenediaminetetraacetic acid disodium[J].Journal of Enzyme Inhibition,2001,16(4):331-319[21] Janna B B,Das S K.Bioturbation induced changes of fertilizer value of phosphate rock in retation to alkaline phosphatase activtity[J].Aquaculture,1992,103(3-4):321-330[22] Newman S.Sediment resuspension effects on alkaline phosphatase activity[J].Hydrobiologia,1992,245:75-86[23] Laura S,Paul I B.Effect of polyphenolic compounds on alkaline phosphatase activity:Its implication for phosphorus regeneralion in Australian freshwaters[J].Arch.Hydrobiol.,1991,123(1):1-19[24] Buainain L B,Kaowaki M K,Maria de Lourdes Polizeli: et al.Characterization of a conidial alkaline phosphatase from the thernophilic fungus Humicola grisea var.thermoidea [J].Basic Microbiol.,1998,38(2):85-94[25] Zhou Y Y, Li J Q,Fu Y Q,et al.Kinetics of alkaline phosphatase in lake sediment associated with cage culture of Oreochromis nilotucus [J].Apuaculture, 2001,203:23-32[26] János Oláh,Erzsébet O.Tth.The function of alkaline phosphatase enzyme in the phosphorus cycle of fertized fishponds [J].Aquacultura Hungarica(Szarvas),1978,1:15-23[27] Barik S K,Purushothaman C S,Mohanty A N.Phosphatase activity with refercnce to bacteria and phosphorus in tropical freshwater aquaculture pond systems[J].Aquaculture Research,2001,32(10):819-832[28] Scholz O,Marxsen J.Sediment phosphatases of the Breitenbach (Germany), a first-order central European stream [J].Arch.Hudrobiol.,1996,135(4):433-450
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