有机碳对海洋着色菌YL28去除无机三态氮的影响
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摘要
【目的】在无机三态氮(氨氮、亚硝氮和硝氮)共存的模拟海水水体中,阐明有机碳尤其是海藻寡糖对海洋着色菌(Marichromatium gracile)YL28生长及去除无机三态氮的影响规律。【方法】采用次溴酸钠氧化法、N-(1-萘基)-乙二胺分光光度法和紫外分光光度法分别测定水体中氨氮、亚硝氮和硝氮的含量,菌体生物量采用比浊法测定。【结果】在光照厌氧环境中,小分子有机酸盐(乙酸钠、丙酮酸钠、琥珀酸钠和柠檬酸钠)是YL28生长和去除无机三态氮的良好有机碳,亚硝氮、硝氮和氨氮去除率分别达到97.92%、99.98%、73.23%-87.15%。单糖(葡萄糖和果糖)、双糖(麦芽糖和蔗糖)和寡糖(壳寡糖和海藻寡糖)是YL28可利用的有机碳,亚硝氮和硝氮去除率分别达到99%和87%以上,氨氮去除率在44.82%-54.53%之间。多糖(β-环糊精、淀粉、黄原胶、琼脂粉、海藻酸钠和卡拉胶)不是菌体利用和去除无机三态氮的有机碳。酵母提取物可作为菌体生长、去除硝氮和亚硝氮的良好有机碳,但严重抑制氨氮去除。海藻酸钠、β-环糊精和卡拉胶分别与乙酸钠共存时,YL28生长和对无机三态氮去除能力与乙酸钠为唯一有机碳的水平相当。乙酸钠体系中添加海藻寡糖,YL28生长速率、最大生物量以及氨氮的去除速率和最大去除率均升高,添加酵母提取物时,生长速率和最大生物量升高,但氨氮去除速率和最大去除率降低。黑暗厌氧环境下,以乙酸钠和氨氮为唯一有机碳和氮源时,YL28不生长,但在无机三态氮共存时,则能良好生长并去除无机三态氮。【结论】在无机三态氮共存海水体系和厌氧条件下,无论是光照还是黑暗环境,YL28均能良好地生长和去除无机三态氮,小分子有机酸盐(乙酸钠、丙酮酸钠、琥珀酸钠、柠檬酸钠)是其良好的有机碳,相对而言,乙酸钠和丙酮酸钠更好。海藻寡糖与乙酸钠复合可提高菌体生长和脱氮能力。本研究为研制开发高效脱氮微生物制剂及其合理性应用提供了指导。
[Objective] Inorganic nitrogens(ammonium, nitrate and nitrite) existed simultaneously in complicated mariculture water. This work aims to explore the effects of organic carbons, especially seaweed oligosaccharides on the removal of inorganic nitrogens by a marine purple sulfur bacterium Marichramatium gracile YL28, which is capable of growth on nitrite as sole nitrogen source. [Methods] Sodium hypobromite oxidation, N-(1-naphthyl)-1,2-diaminoethane dihydrochloride spectrophotometry, and UV spectrophotometry were used for the determination of ammonia, nitrite and nitrate, respectively. Biomass was measured by turbidimetry. [Results] Under anaerobic light condition, organic acid salts(acetate, pyruvate, succinate, citrate) were better carbon sources for YL28, and the removal rate of ammonia, nitrate and nitrite reached 97.92%, 99.98%, and 73.23%-87.15%, respectively. Monosaccharides(glucose and fructose), disaccharides(sucrose and maltose) and oligosaccharides(chito-oligosaccharide and seaweed oligosaccharides) were also suitable for YL28 growth. The remove rate of nitrite and nitrate reached more than 99% and 87%, respectively. While the remove rate of ammonia was 44.82%-54.53%. YL28 had poor growth when polysaccharides(alginate, β-cyclodextrin, starch, xanthan gum, carrageenan and agar) as carbon sources. Yeast extract is favorable for YL28 growth, but it severely inhibited the removal of ammonia. Interestingly, the cell growth and inorganic nitrogen removal in combination of poor and good carbon source systems was equally well to that in good carbon source system. The combination of seaweed oligosaccharides and acetate sodium or yeast extract and acetate sodium promoted the increasing growth rate and biomass, however, there was a significantly different in removal rate of ammonium, seaweed oligosaccharides promoted the removal rate of ammonia, while yeast extract not. Under anaerobic dark condition, when using sodium acetate and ammonia as sole carbon or nitrogen source, YL28 grew poorly. However, under coexisting inorganic nitrogen environment, YL28 not only grew well but also removed efficiently inorganic nitrogen. [Conclusion] In coexisting inorganic nitrogen environment, whatever anaerobically in the light or dark, YL28 grows well and has better removal capacity to inorganic nitrogen. Organic acid salts are better carbon sources, sodium acetate is most suitable carbon source. The combination of seaweed oligosaccharides and sodium acetate could significantly promote cell growth and removal of inorganic nitrogen. This study provides valuable information for the development of higher efficiency water cleaner for sustainable mariculture.
[Objective] Inorganic nitrogens(ammonium, nitrate and nitrite) existed simultaneously in complicated mariculture water. This work aims to explore the effects of organic carbons, especially seaweed oligosaccharides on the removal of inorganic nitrogens by a marine purple sulfur bacterium Marichramatium gracile YL28, which is capable of growth on nitrite as sole nitrogen source. [Methods] Sodium hypobromite oxidation, N-(1-naphthyl)-1,2-diaminoethane dihydrochloride spectrophotometry, and UV spectrophotometry were used for the determination of ammonia, nitrite and nitrate, respectively. Biomass was measured by turbidimetry. [Results] Under anaerobic light condition, organic acid salts(acetate, pyruvate, succinate, citrate) were better carbon sources for YL28, and the removal rate of ammonia, nitrate and nitrite reached 97.92%, 99.98%, and 73.23%-87.15%, respectively. Monosaccharides(glucose and fructose), disaccharides(sucrose and maltose) and oligosaccharides(chito-oligosaccharide and seaweed oligosaccharides) were also suitable for YL28 growth. The remove rate of nitrite and nitrate reached more than 99% and 87%, respectively. While the remove rate of ammonia was 44.82%-54.53%. YL28 had poor growth when polysaccharides(alginate, β-cyclodextrin, starch, xanthan gum, carrageenan and agar) as carbon sources. Yeast extract is favorable for YL28 growth, but it severely inhibited the removal of ammonia. Interestingly, the cell growth and inorganic nitrogen removal in combination of poor and good carbon source systems was equally well to that in good carbon source system. The combination of seaweed oligosaccharides and acetate sodium or yeast extract and acetate sodium promoted the increasing growth rate and biomass, however, there was a significantly different in removal rate of ammonium, seaweed oligosaccharides promoted the removal rate of ammonia, while yeast extract not. Under anaerobic dark condition, when using sodium acetate and ammonia as sole carbon or nitrogen source, YL28 grew poorly. However, under coexisting inorganic nitrogen environment, YL28 not only grew well but also removed efficiently inorganic nitrogen. [Conclusion] In coexisting inorganic nitrogen environment, whatever anaerobically in the light or dark, YL28 grows well and has better removal capacity to inorganic nitrogen. Organic acid salts are better carbon sources, sodium acetate is most suitable carbon source. The combination of seaweed oligosaccharides and sodium acetate could significantly promote cell growth and removal of inorganic nitrogen. This study provides valuable information for the development of higher efficiency water cleaner for sustainable mariculture.
引文
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[12]Zhao CG,Zhang Y,Chan ZH,et al.Insights into arsenic multi-operons expression and resistance mechanisms in Rhodopseudomonas palustris CGA009[J].Frontiers in Microbiology,2015,6:986
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[16]Zheng H,Zhang DM,Wang YN,et al.Removal of inorganic nitrogen and COD from carp farming waste water byRhodopseudomonas[J].Ecological Science,2012,31(4):435-440(in Chinese)郑卉,张德民,王一农,等.红假单胞菌去除养鱼废水三态氮及COD的研究[J].生态科学,2012,31(4):435-440
[17]Jiang P,Hong X,Zhao CG,et al.Reciprocal transformation of inorganic nitrogen by resting cells of Marichromatium gracile YL28[J].Journal of Huaqiao University(Natural Science),2015,36(2):185-189(in Chinese)蒋鹏,洪璇,赵春贵,等.海洋着色菌Marichromatium gracileYL28静息细胞对无机三态氮的相互转化作用[J].华侨大学学报:自然科学版,2015,36(2):185-189
[18]Jiang P,Zhao CG,Yang SP.Influences of low-molecular-weight organic carbon and nitrogen sources on growth and inorganic nitrogen removal by Marichromatium gracile strain YL28[J].Oceanologia et Limnologia Sinica,2014,45(6):1218-1224(in Chinese)蒋鹏,赵春贵,杨素萍.小分子有机碳、氮源对海洋着色菌(Marichromatium gracile)生长和去除高浓度无机三态氮的影响[J].海洋与湖沼,2014,45(6):1218-1224
[19]Jiang P,Zhao CG,Jia YQ,et al.Effects of nitrite on ammonia-nitrogen removal and nitrite-nitrogen as well as photopigment biosynthesis of Marichromatium gracile YL28[J].Microbiology China,2015,42(7):1216-1223(in Chinese)蒋鹏,赵春贵,贾雅琼,等.亚硝氮对海洋着色菌亚硝氮和氨氮去除以及光合色素合成的影响[J].微生物学通报,2015,42(7):1216-1223
[20]Zhao JY,Fu YN,Zhao CG,et al.Identification and characterization of a purple sulfur bacterium from mangrove with rhodopin as predominant carotenoid[J].Acta Microbiologica Sinica,2011,51(10):1318-1325(in Chinese)赵江艳,傅英楠,赵春贵,等.一株高含玫红品的红树林海洋紫色硫细菌分离鉴定及特性[J].微生物学报,2011,51(10):1318-1325
[21]Dang HY,Jiao NZ.Perspectives on the microbial carbon pump with special reference to microbial respiration and ecosystem efficiency in large estuarine systems[J].Biogeosciences,2014,11(14):3887-3898
[22]Jiao NZ,Legendre L,Robinson C,et al.Comment on“Dilution limits dissolved organic carbon utilization in the deep ocean”[J].Science,2015,350(6267):1483
[23]Hou YP,Gao JL,Gu L,et al.Effects of agaro-oligosaccharide treatment on postharvest quality of cherry tomatoes during cold storage[J].Journal of Food Processing and Preservation,2015:39(6):949-955
[24]Zhang ZX,Xu JC,Sheng T,et al.Effect of alginate-derived oligosaccharide on the rape(Brassica chinensis L.)and soil contaminated with cadmium or lead[J].Science and Technology of Food Industry,2014,35(7):49-52,56(in Chinese)张朝霞,许加超,盛泰,等.海藻寡糖对镉、铅单一污染的小油菜及土壤的影响[J].食品工业科技,2014,35(7):49-52,56
[25]Tang JC,Wang F,Chu HR,et al.Bioremediation of petroleum contaminated soils by combination of Zea Mexicana and alginate oligosaccharides[J].Journal of Agro-Environment Science,2010,29(11):2107-2113(in Chinese)唐景春,王斐,褚洪蕊,等.玉米草(Zea Mexicana)与海藻寡糖联合修复石油烃污染土壤的研究[J].农业环境科学学报,2010,29(11):2107-2113
[26]Foss A,Imsland AK,Roth B,et al.Effects of chronic and periodic exposure to ammonia on growth and blood physiology in juvenile turbot(Scophthalmus maximus)[J].Aquaculture,2009,296(1/2):45-50
[27]Gutierrez-Wing MT,Malone RF.Biological filters in aquaculture:trends and research directions for freshwater and marine applications[J].Aquacultural Engineering,2006,34(3):163-171
[28]Liu SJ,Sun Y,Cen YH,et al.Biological control of nitrite in fish farming ponds by Photosynthetic nonsulfer bacteria[J].Environmental Science,1995,16(6):21-23(in Chinese)刘双江,孙燕,岑运华,等.采用光合细菌控制水体中亚硝酸盐的研究[J].环境科学,1995,16(6):21-23
[29]Hou Y,Sun JD,Xu JQ,et al.Degrading characters of ammonia nitrogen in aquatic water of Bacillus megaterium[J].Journal of Shenyang Agricultural University,2006,37(4):607-610(in Chinese)侯颖,孙军德,徐建强,等.巨大芽孢杆菌对养殖水体氨氮降解特性研究[J].沈阳农业大学学报,2006,37(4):607-610
[2]Tang WZ,Shan BQ,Cui JG,et al.Effects of nitrogen pollution on periphyton distribution,elemental composition and assemblage shifts in river ecosystems[J].Clean-Soil Air Water,2015,43(10):1375-1380
[3]Crab R,Avnimelech Y,Defoirdt T,et al.Nitrogen removal techniques in aquaculture for a sustainable production[J].Aquaculture,2007,270(1/4):1-14
[4]He RQ.Environmental remediation a key issue in microbiology China[J].Microbiology China,2012,39(1):1(in Chinese)赫荣乔.环境修复成为我国微生物研究的热点领域[J].微生物学通报,2012,39(1):1
[5]Qi ZZ,Zhang XH,Boon N,et al.Probiotics in aquaculture of China-current state,problems and prospect[J].Aquaculture,2009,290(1/2):15-21
[6]Newaj-Fyzul A,Al-Harbi AH,Austin B.Review:developments in the use of probiotics for disease control in aquaculture[J].Aquaculture,2014,431:1-11
[7]Jiang P,Zhao CG,Jia YQ,et al.Inorganic nitrogen removal by a marine purple sulfur bacterium capable of growth on nitrite as sole nitrogen source[J].Microbiology China,2014,41(5):824-831(in Chinese)蒋鹏,赵春贵,贾雅琼,等.以亚硝氮为唯一氮源生长的海洋紫色硫细菌去除无机三态氮[J].微生物学通报,2014,41(5):824-831
[8]Zhou YB,Liu YJ.Primary analysis of the energy budget and flow in the shrimp pond ecosystem[J].Acta Ecologica Sinica,2000,20(3):474-481(in Chinese)周一兵,刘亚军.虾池生态系能量收支和流动的初步分析[J].生态学报,2000,20(3):474-481
[9]Statton J,Cambridge ML,Dixon KW,et al.Aquaculture of Posidonia australis seedlings for seagrass restoration programs:effect of sediment type and organic enrichment on growth[J].Restoration Ecology,2013,21(2):250-259
[10]Li G,Wu ZB,Hou YS,et al.Nitrogen biotransformation and its relative microorganism in aquaculture waters[J].Chinese Journal of Eco-Agriculture,2006,14(1):11-15(in Chinese)李谷,吴振斌,侯燕松,等.养殖水体氮的生物转化及其相关微生物研究进展[J].中国生态农业学报,2006,14(1):11-15
[11]Harwood CS.Degradation of aromatic compounds by purple nonsulfur bacteria[A]//Hunter CN,Daldal F,Thurnauer MC,et al,eds.The Purple Phototrophic Bacteria[M].Berlin:Springer Netherlands,2009:577-594
[12]Zhao CG,Zhang Y,Chan ZH,et al.Insights into arsenic multi-operons expression and resistance mechanisms in Rhodopseudomonas palustris CGA009[J].Frontiers in Microbiology,2015,6:986
[13]Zhao L,Zhao CG,Han DX,et al.Anaerobic utilization of phenanthrene by Rhodopseudomonas palustris[J].Biotechnology Letters,2011,33(11):2135-2140
[14]Chen YH,Yang ZH,Yu GH,et al.Influence of illumination,oxygen,p H and salinity on growth and nitrite removal effect of Rhodopseudomonas palustris strain 2-8[J].South China Fisheries Science,2010,6(4):1-5(in Chinese)陈燕红,杨紫红,喻国辉,等.光照、氧气、p H和盐度对沼泽红假单胞菌2-8生长和亚硝酸盐消除的影响[J].南方水产,2010,6(4):1-5
[15]Yu JA,Zhang CK,Chen F,et al.Denitration of photosynthetic bacteria P4 strain[J].Journal of Shanghai Jiaotong University,2000,34(11):1579-1582,1597(in Chinese)俞吉安,张承康,陈峰,等.光合细菌P4株的反硝化作用[J].上海交通大学学报,2000,34(11):1579-1582,1597
[16]Zheng H,Zhang DM,Wang YN,et al.Removal of inorganic nitrogen and COD from carp farming waste water byRhodopseudomonas[J].Ecological Science,2012,31(4):435-440(in Chinese)郑卉,张德民,王一农,等.红假单胞菌去除养鱼废水三态氮及COD的研究[J].生态科学,2012,31(4):435-440
[17]Jiang P,Hong X,Zhao CG,et al.Reciprocal transformation of inorganic nitrogen by resting cells of Marichromatium gracile YL28[J].Journal of Huaqiao University(Natural Science),2015,36(2):185-189(in Chinese)蒋鹏,洪璇,赵春贵,等.海洋着色菌Marichromatium gracileYL28静息细胞对无机三态氮的相互转化作用[J].华侨大学学报:自然科学版,2015,36(2):185-189
[18]Jiang P,Zhao CG,Yang SP.Influences of low-molecular-weight organic carbon and nitrogen sources on growth and inorganic nitrogen removal by Marichromatium gracile strain YL28[J].Oceanologia et Limnologia Sinica,2014,45(6):1218-1224(in Chinese)蒋鹏,赵春贵,杨素萍.小分子有机碳、氮源对海洋着色菌(Marichromatium gracile)生长和去除高浓度无机三态氮的影响[J].海洋与湖沼,2014,45(6):1218-1224
[19]Jiang P,Zhao CG,Jia YQ,et al.Effects of nitrite on ammonia-nitrogen removal and nitrite-nitrogen as well as photopigment biosynthesis of Marichromatium gracile YL28[J].Microbiology China,2015,42(7):1216-1223(in Chinese)蒋鹏,赵春贵,贾雅琼,等.亚硝氮对海洋着色菌亚硝氮和氨氮去除以及光合色素合成的影响[J].微生物学通报,2015,42(7):1216-1223
[20]Zhao JY,Fu YN,Zhao CG,et al.Identification and characterization of a purple sulfur bacterium from mangrove with rhodopin as predominant carotenoid[J].Acta Microbiologica Sinica,2011,51(10):1318-1325(in Chinese)赵江艳,傅英楠,赵春贵,等.一株高含玫红品的红树林海洋紫色硫细菌分离鉴定及特性[J].微生物学报,2011,51(10):1318-1325
[21]Dang HY,Jiao NZ.Perspectives on the microbial carbon pump with special reference to microbial respiration and ecosystem efficiency in large estuarine systems[J].Biogeosciences,2014,11(14):3887-3898
[22]Jiao NZ,Legendre L,Robinson C,et al.Comment on“Dilution limits dissolved organic carbon utilization in the deep ocean”[J].Science,2015,350(6267):1483
[23]Hou YP,Gao JL,Gu L,et al.Effects of agaro-oligosaccharide treatment on postharvest quality of cherry tomatoes during cold storage[J].Journal of Food Processing and Preservation,2015:39(6):949-955
[24]Zhang ZX,Xu JC,Sheng T,et al.Effect of alginate-derived oligosaccharide on the rape(Brassica chinensis L.)and soil contaminated with cadmium or lead[J].Science and Technology of Food Industry,2014,35(7):49-52,56(in Chinese)张朝霞,许加超,盛泰,等.海藻寡糖对镉、铅单一污染的小油菜及土壤的影响[J].食品工业科技,2014,35(7):49-52,56
[25]Tang JC,Wang F,Chu HR,et al.Bioremediation of petroleum contaminated soils by combination of Zea Mexicana and alginate oligosaccharides[J].Journal of Agro-Environment Science,2010,29(11):2107-2113(in Chinese)唐景春,王斐,褚洪蕊,等.玉米草(Zea Mexicana)与海藻寡糖联合修复石油烃污染土壤的研究[J].农业环境科学学报,2010,29(11):2107-2113
[26]Foss A,Imsland AK,Roth B,et al.Effects of chronic and periodic exposure to ammonia on growth and blood physiology in juvenile turbot(Scophthalmus maximus)[J].Aquaculture,2009,296(1/2):45-50
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