菹草内生硫氧化细菌的分离鉴定及对鸡粪中硫化氢的减释作用
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摘要
鸡粪中的含硫有机物在微生物作用下会释放出大量的硫化氢气体,危害畜禽的健康及污染环境。试验以减少鸡粪中硫化氢释放量为目的,从菹草内分离得到1株具有异养硫氧化作用的细菌,命名为JS3。16S rRNA序列分析表明菌株JS3与杀鲑气单胞菌的亲源关系最近。以硫代硫酸钠为唯一硫源时菌株JS3的硫氧化特征:碳源优先利用顺序为葡萄糖>柠檬酸钠>乙酸钠;氮源优先利用顺序为硝酸钠>氯化铵;菌株JS3氧化硫的最佳碳氮比为15∶1,此时77.82%的硫代硫酸根被转化为硫酸根;pH从4.05增加到10.05,菌株的氧化硫能力先增强后下降,当pH为9.05时,硫酸根的积累量最高,可达到4.33 g/L;当氯化钠添加量达到4.0%时对菌株JS3才产生明显影响,说明株菌具有一定的耐盐性;最佳硫代硫酸根初始浓度为2 810 mg/L,此时83.22%的硫代硫酸根被转化为硫酸根;250 mL三角瓶中装液量为75 mL时,菌株JS3是硫氧化能力最强。在硫化氢减释模拟试验中,与对照相比,菌株JS3可降低84.53%的硫化氢释放量。
Sulfur-containing organic matter in chicken manure under the action of microorganisms can release a large amount of hydrogen sulfide gas,which can endanger health of livestock and pollute the environment. A strain of endophyte named JS3 with the capacity of heterotrophic sulfur oxidation was isolated from Potamogeton crispus for the purpose of reducing the release of H2 S in chicken manure. 16 S rRNA sequence analysis showed that strain JS3 had the closest relationship with Aeromonas salmonicida. The sulfur oxidation characteristics of strain JS3 with sodium thiosulfate as sole sulfur source were as follows: priority utilization order of glucose > sodium citrate > sodium acetate;priority utilization order of strain JS3 to oxidize sulfur was ammonium nitrate > ammonium chloride;77. 82% of thiosulfate was converted to sulfate with the best ratio of carbon to nitrogen of 15 ∶ 1;sulfur oxidation ability of strain JS3 increased and then decreased,when pH increased from 4. 05 to10. 05 and the accumulation of sulfate was up to 4. 33 g/L with the optimal pH of 9. 05; sulfur oxidation ability of strain JS3 was significantly affected when additive amount of sodium chloride was up to 4%,which indicated that strain JS3 had high salt tolerance; the optimal initial concentration of thiosulfate for strain JS3 to oxidize sulfur was 2 810 mg/L,with 83. 22% of thiosulfate being converted to sulfate; the best liquid volume was 75 mL in 250 mL triangular bottle for strain JS3 to oxidize sulfur with 80. 72% of thiosulfate being converted to sulfate. In simulation experiment,strain JS3 could reduce 84. 53% of the amount of released H2 S compared with the control group.
Sulfur-containing organic matter in chicken manure under the action of microorganisms can release a large amount of hydrogen sulfide gas,which can endanger health of livestock and pollute the environment. A strain of endophyte named JS3 with the capacity of heterotrophic sulfur oxidation was isolated from Potamogeton crispus for the purpose of reducing the release of H2 S in chicken manure. 16 S rRNA sequence analysis showed that strain JS3 had the closest relationship with Aeromonas salmonicida. The sulfur oxidation characteristics of strain JS3 with sodium thiosulfate as sole sulfur source were as follows: priority utilization order of glucose > sodium citrate > sodium acetate;priority utilization order of strain JS3 to oxidize sulfur was ammonium nitrate > ammonium chloride;77. 82% of thiosulfate was converted to sulfate with the best ratio of carbon to nitrogen of 15 ∶ 1;sulfur oxidation ability of strain JS3 increased and then decreased,when pH increased from 4. 05 to10. 05 and the accumulation of sulfate was up to 4. 33 g/L with the optimal pH of 9. 05; sulfur oxidation ability of strain JS3 was significantly affected when additive amount of sodium chloride was up to 4%,which indicated that strain JS3 had high salt tolerance; the optimal initial concentration of thiosulfate for strain JS3 to oxidize sulfur was 2 810 mg/L,with 83. 22% of thiosulfate being converted to sulfate; the best liquid volume was 75 mL in 250 mL triangular bottle for strain JS3 to oxidize sulfur with 80. 72% of thiosulfate being converted to sulfate. In simulation experiment,strain JS3 could reduce 84. 53% of the amount of released H2 S compared with the control group.
引文
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[24]王思珍,东彦新,夏春丽,等.鸡舍内有害气体及调控措施[J].内蒙古民族大学学报(自然科学版),2002,17(1):48-51.
[2]李婉,杨承剑,梁贤威,等.畜禽粪便恶臭气体产生与调控的微生物学研究进展[J].中国畜牧兽医,2013,40(8):195-198.
[3]李明堂,林泳墨.畜禽养殖排放内源类固醇雌激素的污染与控制研究进展[J].吉林农业大学学报,2014,36(3):253-257,264.
[4]Zicari M K.Removal of hydrogen sulfide from biogas using cow-manure compost[D].New York:Graduate School of Cornell University,2003.
[5]刘春梅,徐凤花,曹艳花,等.除臭菌株对NH3和H2S释放及物质转化的影响[J].农业环境科学学报,2011,30(3):585-590.
[6]朱淑斌.除臭剂在养猪生产中的应用[J].湖北畜牧兽医,2008(6):12-14.
[7]Chou S,Mike F,Sam K,et al.Toxicological profile for hydrogen sulfide[M].Atlanta:US Public Health Services Agency for Toxic Substances and Disease Registry,2006:143-152.
[8]Luc M,Catherine G,Jean-Claude R,et al.Biological treatment process of air loaded with an ammonia and hydrogen sulfide mixture[J].Chemosphere,2003,50(1):145-153.
[9]梁美生,苏娟.脱除养殖臭气中硫化氢菌种的筛选及性能测定[J].中国矿业大学学报,2012,41(5):844-849.
[10]李彪,熊焰.猪粪中除臭微生物的筛选和鉴定[J].家畜生态学报,2008,29(1):74-76.
[11]李莎,蒋文举,金燕,等.微生物脱臭研究应用进展[J].环境科学与技术,2007,30(5):108-110.
[12]Syed M,Soreanu G,Falletta P,et al.Removal of hydrogen sulfide from gas streams using biological processes-a review[J].Canadian Biosystems Engineering,2006,48:21-24.
[13]Gommers P J F,Buleveld W,Zuijderwijk F J M,et al.Simultaneous sulfide and acetate in a denitrifying fluidized bed reactor and reactor performance[J].Water Research,1988(22):1075-1083.
[14]Kanagawa T,Mikami E.Removal of methanethiol dimethyl sulfide dimethyl disulfide and hydrogen sulfide from contaminated air by thiobacillus thioparus TK-m[J].Applied and Environmental Microbiology,1989,55(3):555-558.
[15]徐桂芹,姜安玺,张英民,等.假单胞菌固定化除硫化氢臭气的试验研究[J].给水排水,2005,31(2):41-43.
[16]Palleroni N J,Krieg N R,Holt J G.Bergey's Manual of Systematic Bacteriology[M].Beijing:The Press of Science,1984:443-491.
[17]东秀株,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:1-15.
[18]陈庆丽,景澄茗,付韵馨,等.寒区水体中溶藻铜绿假单胞菌的分离和性质研究[J].环境科学学报,2015,35(3):692-698.
[19]王孝镕.硫离子、亚硫酸根和硫代硫酸根的分光光度测定[J].烟台师院学报,1986,2(2):18-22.
[20]朱国英,管健.离子色谱法和铬酸钡分光光度法测定水中硫酸盐的比较[J].环境与健康杂志,2008,25(2):171.
[21]宋艳晶.鸡粪产臭气微生物洁抗菌的分离与鉴定[D].保定:河北农业大学,2009.
[22]龙金满.亚甲基蓝比色法测定大气中的硫化氢[J].云南环境科学,2003(41):34-42.
[23]王锐,邱丽娜,关景洋,等.一株嗜酸硫氧化细菌的分离鉴定及生长特性[J].环境科学学报,2010,30(3):485-489.
[24]王思珍,东彦新,夏春丽,等.鸡舍内有害气体及调控措施[J].内蒙古民族大学学报(自然科学版),2002,17(1):48-51.