藻菌共生生物膜系统修复煤炭矿区污染水体的研究
摘要
煤炭作为中国最主要的能源之一,对我国社会经济有着极其重要的影响,但在煤炭的开采和利用过程中对水环境产生了巨大的危害。在赴陕北神木煤炭矿区实地调查和采样分析的基础上,采用单因子水质标识指数和综合水质标识指数对4处水样进行评价,结果显示：陕北煤炭矿区水体受煤粉污染较为严重,水体中色度、悬浮物、有机物、NH4+-N、NO3-、SO42、Cr6+、Cu2+、Fe3+、Mn2+、Pb、Cd元素比参照水样都有不同程度的升高,尤其是重金属Cr和Cd、COD和TP,都达到严重污染环境的水平；六道沟煤矸石淋溶形成的淤泥附近水样污染最为严重,主要超标污染物为悬浮物、磷酸盐、Cr6+、COD和Cd,综合水质标识指数为3.561。
针对煤炭矿区污染水体的特点和陕北光照充足的优势,采取藻菌共生生物膜系统修复这一污染水体,利用球形填料和弹性立体填料作为生物附着生长的载体,以Cr(Ⅵ)作为代表重金属,在推流式氧化塘上架设日光灯管模拟自然光照,光暗比14：10,考察了藻菌共生生物膜对Cr(Ⅵ)、COD、NH4+-N、TP、SO42-的去除效果和去除机理,得到以下结论：
(1)从氧化沟活性污泥中分离筛选出6株耐铬菌株,发现菌株在培养24h后生长状况最好；以耐铬性最好的2-5号菌株做试验,结果显示菌株对Cr(Ⅵ)的去除效率随初始Cr(Ⅵ)浓度的升高呈先上升后下降的趋势,具有最优去除浓度区间和相应的最大去除效率；在含Cr(Ⅵ)浓度为654mg/L的废水中培养24h后,2-5号菌株Cr(Ⅵ)的去除率最高可达76.4%,去除速率为18.9mg/(gSS·h);当废水中含Cr(Ⅵ)浓度较低时,菌废比较高有利于增大去除率,当Cr(Ⅵ)浓度较高时,增大菌废比对去除率影响不大；6株耐铬菌均有较好的生物累积Cr(Ⅵ)的能力,但耐Cr(Ⅵ)能力与去除Cr(Ⅵ)的能力之间没有直接关系。
(2)在HRT为14.3d,载体为球状填料时,藻菌共生生物膜系统对Cr(Ⅵ)有较高的去除率,最高可达98.3%,能忍耐21.2mg/LCr(Ⅵ),此时去除率为88.2%；在进水端底泥中Cr(Ⅵ)含量高达286μg/g干重底泥,Cr(Ⅵ)主要是被生物还原为Cr(Ⅲ),然后生成Cr(OH)3沉淀进入底泥中；水绵对Cr(Ⅵ)的吸附及积累能力比凤眼莲低,水绵中Cr(Ⅵ)含量为12.08μg/g干重,凤眼莲为79.41μg/g干重。
(3)COD进水浓度为100mg/L时,去除率稳定在80%以上,利用光照氧化塘可以实现异养细菌和光合生物的互惠共生,协同去除重金属和有机物;NH4+-N进水浓度为6mg/L左右时,最高去除率达65.3%,主要依靠同化作用和硝化去除,夏季升温和水绵的添加有助于NH4+-N去除率的提高,NH4+-N在氧化塘前半部分约有87%被去除；进水中TP为1.7mg/L左右时,TP去除率逐渐稳定在60%以上,最高达76.2%。氧化塘前半部分的去除率约占总去除率的60%,主要依靠同化作用及生成沉淀从氧化塘中去除。
(4)进水COD为50mg/L时,光照氧化塘内昼间由于藻类光合产氧,溶解氧平均浓度为3.5mg/L,不具备SO42-厌氧还原的条件,SO42-去除率低于10%。进水COD为100mg/L时,昼间溶解氧为1.8mg/L,晚上8：00熄灯时溶解氧最低为0.72mg/L,熄灯一小时后溶解氧最低0.37mg/L,可实现SO42-厌氧还原,在进水SO42-浓度为70.7mg/L时,最大去除率可达65.5%。平均有82%的SO42-在氧化塘的前半部分去除。
(5)藻菌共生生物膜系统从启动到稳定运行,光照氧化塘内生物大致经历了3个大的变化阶段：水生植物阶段、凤眼莲阶段和水绵生物膜阶段。光照不足、底泥较浅及COD耗氧形成厌氧环境都易导致水生植物死亡；凤眼莲阶段氧化塘内生物多样性丰富,包括水蜗牛、毛饰拟剑水蚤等动物及多种微藻,微藻以绿藻为主；光照强度增大有助于增加水蜗牛及毛饰拟剑水蚤的数量；凤眼莲对周围微生物形成抑制,并可阻止水绵在载体上挂膜；进水Cr(Ⅵ)浓度达10mg/L,氧化塘内Cr(Ⅵ)浓度为1.6mg/L时,凤眼莲无法耐受而死亡；凤眼莲抑制解除后水绵可在载体上形成生物膜,水绵根部多附着在细菌上生长,靠细菌分泌的胞外多聚物与载体粘连；水绵生物膜上藻丝长度(μm)与时间(d)大致关系为y=48.77x,水绵生物膜阶段微藻以蓝藻为主；在485nm处监测4个采样点处的OD值,结果显示进水污染物浓度较高、溶解氧浓度较低时悬浮微藻的数量较小,光照作用强烈时水中悬浮微藻数量增加。
(6)水绵在立体弹性载体的细丝上缠绕生长,藻丝较长,形成的生物膜较薄(300-500μm),光照及水深对藻类的数量有决定性的影响,细菌的数量主要由COD和Cr(Ⅵ)浓度决定；增加立体弹性载体可增加氧化塘上部的藻类浓度和氧化塘上部、下部的细菌浓度,有助于强化缺氧及厌氧的环境；增加立体弹性载体对COD和TP的去除帮助不大,对Cr(Ⅵ)的去除以及硝化和反硝化作用有改善作用,对SO42-的去除有较好的促进作用。
(7)HRT的缩短对Cr(Ⅵ)的去除造成的影响较小,HRT为7.2d时,Cr(Ⅵ)去除速率最高可达2.4mg/(L·d)；系统对COD的最大去除能力为12.5mg/(L·d),进一步缩小HRT已不能增大COD的去除负荷；减小HRT使硝化作用受到了Cr(Ⅵ)毒性的抑制,造成NH4+-N去除速率的下降；TP的去除负荷最高仅为0.11mg/(L·d),增加负荷后TP的去除途径彻底被破坏,在Cr(Ⅵ)浓度为20mg/L时,要保证TP有一定的去除率,必须保证HRT在14.3d以上；SRB在3.1mg/(L·d)的Cr(Ⅵ)负荷下依然可以生存良好,并能保证一定的SO42-去除率,去除负荷可达7.5mg/(L·d)。
(8)投加耐铬菌株提高Cr(Ⅵ)的去除效率有短暂帮助,在突发性环境污染事故中可以采用投加高效降解菌株的方法降低污染程度,对于一个连续处理污染物的系统,投加高效降解菌株作用甚小。
Coal is one of the most important energy in China which has a significant influence on our social and economic development. However, coal mining and utilization process caused a serious harmness to local water. Based on pollution investigation and monitoring of4water samples in Shaanbei Shenmu coal mine, the water quality in Shaanbei coal mine were evaluted by methods of single factor water quality identification index and comprehensive water quality identification index. The results showed:coal powder pollution was much serious and color suspended solid (SS), COD, NH4+-N、NO3-,SO42-,Cr6+,Cu2+, Fe3+, Mn2+, Pb. Cd elements in waters were much more than the reference sample, especially heavy metals Cr. Cd. COD and TP were high enough to pollute envrionmental seriously. The water quality near mud caused by coal gangue in Ludaogou was the worst. Its pollutants exceeding the standard were SS. PO43-, COD, Cr, Cd and its comprehensive water quality identification index was3.561.
According to the characteristics of polluted water in coal mining area and the advantage of Shaanbei with sufficient sunlight, symbiotic algae-bacteria biofilm system was employed to remediate this polluted water. The carriers for attachment of microorganisms and algae included spherical packing and elastic three-dimensional packing. Cr (Ⅵ) was hired as a representative of diverse heavy metals. To simulate natural light, three fluorescent tubes were added on the oxidation ponds with a light:dark than14:10. The removal efficiency and removing mechanism of Cr(Ⅵ), COD, NH4+-N. TP, SO4-2-by symbiotic algae-bacteria biofilm were examined. The results was presented as following:
(1) Six Cr-tolerant strains were isolated from the active sludge in a oxidation ditch to test their cellular growth mass in different Cr(Ⅵ) concentration liquid medium after24h,48h and72h growth. A high Cr-tolerant strain was choosed from them to removal Cr(Ⅵ) in wastewater. The cellular mass of six strain were decreased with increasing initial Cr(Ⅵ) concentration in most cases. The high Cr-tolerant strain had a maximum Cr(Ⅵ) removal efficiency(76.4%) in a best concentration range (about654mg/L) and the highest Cr(Ⅵ) removal rate was18.9mg/(gSS·h) after24h growth. A higher ratio of biomass to amount of Cr(Ⅵ) was benefit to improve the removal of Cr(Ⅵ) when the Cr(Ⅵ) concentration was lower. However, when the Cr(Ⅵ) concentration was higher, it did not help much for improve the removal of Cr(Ⅵ). All of them presented excellent bioaccumulation abilities, however, it seemed that the resistance and the removal potential of heavy metal had no direct connection.
(2) When HRT was14.3d and carrier was spheric packing, the algae-bacteria biofilm had a better removal efficiency of98.3%and was tolerant to21.2mg/L Cr(Ⅵ) with a removal potential of88.2%. Cr(Ⅵ) content reacheed286μg/g (dry weight mud) in the sediment of influent side. Cr(Ⅵ) was reduced to Cr(Ⅲ) by microbes. Cr(Ⅲ) turned into Cr(OH)3and then precipitated into sediment. The bioaccumulation potential of Cr(Ⅵ) by Spirogyra (12.08μg/g dry weight) is far less than Eichhornia crassipes's (79.41μg/g dry weight).
(3) When the concentration of COD in influent was100mg/L. the COD removal efficiency was beyond80%. In oxidation pond the heterotrophic bacteria and algae cooperated with each other to reduce heavy metal and COD. The best removal efficiency of NH4+-N was65.3%with a6mg/L influent concentration. Nitrogen was removed by assimilation anabolism and nitrification. The influent TP concentration was1.7mg/L of which over60%was reduced by assimilation and precipitation.
(4) When the inflow COD was50mg/L, the dissolved oxygen(DO) was3.5mg/L in reactor for photosynthetic oxygenation by algae which was unsuitable for the reduction of SO42-. After increasing the COD concentration to100mg/L, the diurnal average DO was1.8mg/L and minor night DO was0.37mg/L in which sulfate reduction could take place. The max removal was65.5%with a70.7mg/Linfluent sulfate.
(5) Algae-bacteria biofilm system went through three periods from start-up to stable operation:hydrophyte stage, Eichhornia crassipe stage and Spirogyra biofilm stage. Eichhomia crassipe could inhibit the formation of Spirogyra biofilm. Green algae grew better in Eichhornia crassipe stage than Spirogyra biofilm.When Eichhornia crassipe was taken out from the reactor Spirogyra began to attach to carriers. Spirogyra prefer to immobilize on extracellular polymeric substances (ESP) excretived by bacteria. The length of Spirogyra filament had a good linear relationship with time(d):y=48.77x.
(6) Illumination and depth of water were important to algae amount, whereas concentration of COD and Cr(Ⅵ) were key factors for bacterial growth. Addition of the tridimensional elastic carrier could increase the algae amount of upper reactor and the bacteria concentration both upper and lower reactor which was helpful to strengthen anoxic and anaerobic environment for better denitrification and sulfate reduction.
(7) A shorter hydraulic retention time (HRT) had little effect on Cr(Ⅵ) removal. For a7.2h HRT, Cr (Ⅵ) removal rate could run up to2.4mg/(L·d). The system had reached the maximum COD removal capability of12.5mg/(L·d). Less HRT could impact nitrogen removal potential and was destructive to TP removal unless with a longer HRT than14.3d, however, sulfate-reducing bacteria(SRB) could still survive under a Cr(Ⅵ) loading rate of3.1mg/(L·d) and remove7.5mg sulfate per litre within a day.
(8)The dosage of Cr-resistance strains had a impermanency improvement for increasing Cr(Ⅵ) removal efficiency. It could be used in a hazardous waste pollution event rather than in a continuous water treatment svstem.
针对煤炭矿区污染水体的特点和陕北光照充足的优势,采取藻菌共生生物膜系统修复这一污染水体,利用球形填料和弹性立体填料作为生物附着生长的载体,以Cr(Ⅵ)作为代表重金属,在推流式氧化塘上架设日光灯管模拟自然光照,光暗比14：10,考察了藻菌共生生物膜对Cr(Ⅵ)、COD、NH4+-N、TP、SO42-的去除效果和去除机理,得到以下结论：
(1)从氧化沟活性污泥中分离筛选出6株耐铬菌株,发现菌株在培养24h后生长状况最好；以耐铬性最好的2-5号菌株做试验,结果显示菌株对Cr(Ⅵ)的去除效率随初始Cr(Ⅵ)浓度的升高呈先上升后下降的趋势,具有最优去除浓度区间和相应的最大去除效率；在含Cr(Ⅵ)浓度为654mg/L的废水中培养24h后,2-5号菌株Cr(Ⅵ)的去除率最高可达76.4%,去除速率为18.9mg/(gSS·h);当废水中含Cr(Ⅵ)浓度较低时,菌废比较高有利于增大去除率,当Cr(Ⅵ)浓度较高时,增大菌废比对去除率影响不大；6株耐铬菌均有较好的生物累积Cr(Ⅵ)的能力,但耐Cr(Ⅵ)能力与去除Cr(Ⅵ)的能力之间没有直接关系。
(2)在HRT为14.3d,载体为球状填料时,藻菌共生生物膜系统对Cr(Ⅵ)有较高的去除率,最高可达98.3%,能忍耐21.2mg/LCr(Ⅵ),此时去除率为88.2%；在进水端底泥中Cr(Ⅵ)含量高达286μg/g干重底泥,Cr(Ⅵ)主要是被生物还原为Cr(Ⅲ),然后生成Cr(OH)3沉淀进入底泥中；水绵对Cr(Ⅵ)的吸附及积累能力比凤眼莲低,水绵中Cr(Ⅵ)含量为12.08μg/g干重,凤眼莲为79.41μg/g干重。
(3)COD进水浓度为100mg/L时,去除率稳定在80%以上,利用光照氧化塘可以实现异养细菌和光合生物的互惠共生,协同去除重金属和有机物;NH4+-N进水浓度为6mg/L左右时,最高去除率达65.3%,主要依靠同化作用和硝化去除,夏季升温和水绵的添加有助于NH4+-N去除率的提高,NH4+-N在氧化塘前半部分约有87%被去除；进水中TP为1.7mg/L左右时,TP去除率逐渐稳定在60%以上,最高达76.2%。氧化塘前半部分的去除率约占总去除率的60%,主要依靠同化作用及生成沉淀从氧化塘中去除。
(4)进水COD为50mg/L时,光照氧化塘内昼间由于藻类光合产氧,溶解氧平均浓度为3.5mg/L,不具备SO42-厌氧还原的条件,SO42-去除率低于10%。进水COD为100mg/L时,昼间溶解氧为1.8mg/L,晚上8：00熄灯时溶解氧最低为0.72mg/L,熄灯一小时后溶解氧最低0.37mg/L,可实现SO42-厌氧还原,在进水SO42-浓度为70.7mg/L时,最大去除率可达65.5%。平均有82%的SO42-在氧化塘的前半部分去除。
(5)藻菌共生生物膜系统从启动到稳定运行,光照氧化塘内生物大致经历了3个大的变化阶段：水生植物阶段、凤眼莲阶段和水绵生物膜阶段。光照不足、底泥较浅及COD耗氧形成厌氧环境都易导致水生植物死亡；凤眼莲阶段氧化塘内生物多样性丰富,包括水蜗牛、毛饰拟剑水蚤等动物及多种微藻,微藻以绿藻为主；光照强度增大有助于增加水蜗牛及毛饰拟剑水蚤的数量；凤眼莲对周围微生物形成抑制,并可阻止水绵在载体上挂膜；进水Cr(Ⅵ)浓度达10mg/L,氧化塘内Cr(Ⅵ)浓度为1.6mg/L时,凤眼莲无法耐受而死亡；凤眼莲抑制解除后水绵可在载体上形成生物膜,水绵根部多附着在细菌上生长,靠细菌分泌的胞外多聚物与载体粘连；水绵生物膜上藻丝长度(μm)与时间(d)大致关系为y=48.77x,水绵生物膜阶段微藻以蓝藻为主；在485nm处监测4个采样点处的OD值,结果显示进水污染物浓度较高、溶解氧浓度较低时悬浮微藻的数量较小,光照作用强烈时水中悬浮微藻数量增加。
(6)水绵在立体弹性载体的细丝上缠绕生长,藻丝较长,形成的生物膜较薄(300-500μm),光照及水深对藻类的数量有决定性的影响,细菌的数量主要由COD和Cr(Ⅵ)浓度决定；增加立体弹性载体可增加氧化塘上部的藻类浓度和氧化塘上部、下部的细菌浓度,有助于强化缺氧及厌氧的环境；增加立体弹性载体对COD和TP的去除帮助不大,对Cr(Ⅵ)的去除以及硝化和反硝化作用有改善作用,对SO42-的去除有较好的促进作用。
(7)HRT的缩短对Cr(Ⅵ)的去除造成的影响较小,HRT为7.2d时,Cr(Ⅵ)去除速率最高可达2.4mg/(L·d)；系统对COD的最大去除能力为12.5mg/(L·d),进一步缩小HRT已不能增大COD的去除负荷；减小HRT使硝化作用受到了Cr(Ⅵ)毒性的抑制,造成NH4+-N去除速率的下降；TP的去除负荷最高仅为0.11mg/(L·d),增加负荷后TP的去除途径彻底被破坏,在Cr(Ⅵ)浓度为20mg/L时,要保证TP有一定的去除率,必须保证HRT在14.3d以上；SRB在3.1mg/(L·d)的Cr(Ⅵ)负荷下依然可以生存良好,并能保证一定的SO42-去除率,去除负荷可达7.5mg/(L·d)。
(8)投加耐铬菌株提高Cr(Ⅵ)的去除效率有短暂帮助,在突发性环境污染事故中可以采用投加高效降解菌株的方法降低污染程度,对于一个连续处理污染物的系统,投加高效降解菌株作用甚小。
Coal is one of the most important energy in China which has a significant influence on our social and economic development. However, coal mining and utilization process caused a serious harmness to local water. Based on pollution investigation and monitoring of4water samples in Shaanbei Shenmu coal mine, the water quality in Shaanbei coal mine were evaluted by methods of single factor water quality identification index and comprehensive water quality identification index. The results showed:coal powder pollution was much serious and color suspended solid (SS), COD, NH4+-N、NO3-,SO42-,Cr6+,Cu2+, Fe3+, Mn2+, Pb. Cd elements in waters were much more than the reference sample, especially heavy metals Cr. Cd. COD and TP were high enough to pollute envrionmental seriously. The water quality near mud caused by coal gangue in Ludaogou was the worst. Its pollutants exceeding the standard were SS. PO43-, COD, Cr, Cd and its comprehensive water quality identification index was3.561.
According to the characteristics of polluted water in coal mining area and the advantage of Shaanbei with sufficient sunlight, symbiotic algae-bacteria biofilm system was employed to remediate this polluted water. The carriers for attachment of microorganisms and algae included spherical packing and elastic three-dimensional packing. Cr (Ⅵ) was hired as a representative of diverse heavy metals. To simulate natural light, three fluorescent tubes were added on the oxidation ponds with a light:dark than14:10. The removal efficiency and removing mechanism of Cr(Ⅵ), COD, NH4+-N. TP, SO4-2-by symbiotic algae-bacteria biofilm were examined. The results was presented as following:
(1) Six Cr-tolerant strains were isolated from the active sludge in a oxidation ditch to test their cellular growth mass in different Cr(Ⅵ) concentration liquid medium after24h,48h and72h growth. A high Cr-tolerant strain was choosed from them to removal Cr(Ⅵ) in wastewater. The cellular mass of six strain were decreased with increasing initial Cr(Ⅵ) concentration in most cases. The high Cr-tolerant strain had a maximum Cr(Ⅵ) removal efficiency(76.4%) in a best concentration range (about654mg/L) and the highest Cr(Ⅵ) removal rate was18.9mg/(gSS·h) after24h growth. A higher ratio of biomass to amount of Cr(Ⅵ) was benefit to improve the removal of Cr(Ⅵ) when the Cr(Ⅵ) concentration was lower. However, when the Cr(Ⅵ) concentration was higher, it did not help much for improve the removal of Cr(Ⅵ). All of them presented excellent bioaccumulation abilities, however, it seemed that the resistance and the removal potential of heavy metal had no direct connection.
(2) When HRT was14.3d and carrier was spheric packing, the algae-bacteria biofilm had a better removal efficiency of98.3%and was tolerant to21.2mg/L Cr(Ⅵ) with a removal potential of88.2%. Cr(Ⅵ) content reacheed286μg/g (dry weight mud) in the sediment of influent side. Cr(Ⅵ) was reduced to Cr(Ⅲ) by microbes. Cr(Ⅲ) turned into Cr(OH)3and then precipitated into sediment. The bioaccumulation potential of Cr(Ⅵ) by Spirogyra (12.08μg/g dry weight) is far less than Eichhornia crassipes's (79.41μg/g dry weight).
(3) When the concentration of COD in influent was100mg/L. the COD removal efficiency was beyond80%. In oxidation pond the heterotrophic bacteria and algae cooperated with each other to reduce heavy metal and COD. The best removal efficiency of NH4+-N was65.3%with a6mg/L influent concentration. Nitrogen was removed by assimilation anabolism and nitrification. The influent TP concentration was1.7mg/L of which over60%was reduced by assimilation and precipitation.
(4) When the inflow COD was50mg/L, the dissolved oxygen(DO) was3.5mg/L in reactor for photosynthetic oxygenation by algae which was unsuitable for the reduction of SO42-. After increasing the COD concentration to100mg/L, the diurnal average DO was1.8mg/L and minor night DO was0.37mg/L in which sulfate reduction could take place. The max removal was65.5%with a70.7mg/Linfluent sulfate.
(5) Algae-bacteria biofilm system went through three periods from start-up to stable operation:hydrophyte stage, Eichhornia crassipe stage and Spirogyra biofilm stage. Eichhomia crassipe could inhibit the formation of Spirogyra biofilm. Green algae grew better in Eichhornia crassipe stage than Spirogyra biofilm.When Eichhornia crassipe was taken out from the reactor Spirogyra began to attach to carriers. Spirogyra prefer to immobilize on extracellular polymeric substances (ESP) excretived by bacteria. The length of Spirogyra filament had a good linear relationship with time(d):y=48.77x.
(6) Illumination and depth of water were important to algae amount, whereas concentration of COD and Cr(Ⅵ) were key factors for bacterial growth. Addition of the tridimensional elastic carrier could increase the algae amount of upper reactor and the bacteria concentration both upper and lower reactor which was helpful to strengthen anoxic and anaerobic environment for better denitrification and sulfate reduction.
(7) A shorter hydraulic retention time (HRT) had little effect on Cr(Ⅵ) removal. For a7.2h HRT, Cr (Ⅵ) removal rate could run up to2.4mg/(L·d). The system had reached the maximum COD removal capability of12.5mg/(L·d). Less HRT could impact nitrogen removal potential and was destructive to TP removal unless with a longer HRT than14.3d, however, sulfate-reducing bacteria(SRB) could still survive under a Cr(Ⅵ) loading rate of3.1mg/(L·d) and remove7.5mg sulfate per litre within a day.
(8)The dosage of Cr-resistance strains had a impermanency improvement for increasing Cr(Ⅵ) removal efficiency. It could be used in a hazardous waste pollution event rather than in a continuous water treatment svstem.
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