四区一体反应器冬季启动脱氮特性及硝化菌群结构分析

我国分散型污水处理问题亟待解决,研究开发了四区一体式生物膜-活性污泥生物脱氮反应器,采用梯度缩短HRT方法,考察了冬季8~15℃水温环境下的反应器COD、NH+4-N、TN去除效果,同时,采用荧光原位杂交技术(FISH)对种泥及其启动过程中的生物膜硝化菌群(AOB、NOB)结构进行分析,并对反应器运行效果与功能微生物的相关关系进行了探讨.结果表明,在HRT为9.2 h下COD、NH+4-N、TN去除率分别为92.11%、99.21%、61.63%;启动末期生物膜内AOB、NOB数量是种泥的5.82倍、6.14倍,硝化细菌占总菌量由6.12%上升至16.38%,成为生物膜的优势菌群;末期硝化效率由初期78.49%上升至97.52%,NOB数量增长5.61倍,AOB/NOB值优化为1.47,反应器内富集生长的AOB、NOB及合适的AOB/NOB比值是确保硝化出水水质的重要保障.     

常温低基质下碱度和溶解氧对厌氧氨氧化的影响

通过改变进水碱度、光照条件和溶解氧,对以陶粒为填料的常温低基质上向流厌氧氨氧化生物滤柱的脱氮性能进行了研究.借助克隆测序技术对滤柱内的微生物菌群进行了分析.结果表明,水温为20℃±2℃,水力停留时间为3 h条件下,当进水碱度在44~350 mg·L-1时,随着碱度的降低,氨氮去除率从97.2%降至75.6%,总氮去除率从89.7%降至75.1%,而亚硝氮去除率稳定在99.7%,进水碱度为0 mg·L-1时,出水亚硝氮浓度突跃至4.9 mg·L-1.当进水碱度恢复至350 mg·L-1,存在自然光照情况下,氨氮去除率较没有光照时的氨氮去除率降低12.3%.水力停留时间为1.5 h条件下,当进水DO小于3mg·L-1时,平均氨氮去除率和亚硝氮去除率分别为99.7%和100%,平均总氮去除负荷为1.0 kg·(m3·d)-1.16S rRNA克隆测序鉴定结果表明,滤柱下部的厌氧氨氧化菌种类为Candidatus Jettenia asiatica和Candidatus Brocadia sp.,这两种菌对20℃±2℃水温有很好的适应性.     

厌氧氨氧化菌的富集培养与分子鉴定

采用传统培养方法与分子生物学方法相结合,进行了厌氧氨氧化菌的分离培养与分子鉴定.富集培养以2种不同反应器的厌氧氨氧化污泥作为分离源,以亚硝酸盐与铵盐为底物.经过2 a传代培养获得2个培养系,对亚硝酸盐及铵盐具有稳定的去除能力,氨氮去除率为85%左右.通过16S rRNA克隆文库方法对培养系中浮霉菌门微生物群落结构进行了多样性解析,结果表明2个培养系中的厌氧氨氧化菌是同一种微生物,代表序列比对结果与"Kuenenia stuttgartiensis"同源性为99%.采用"K.stuttgartiensis"的特异探针对培养系进行了荧光原位杂交分析(FISH),进一步证实"K.stuttgartiensis"是培养系中的优势厌氧氨氧化菌,占总菌群数的80%～90%.对反应器中"K.stuttgartiensis"的丰度变化进行了FISH跟踪检测,发现原始接种污泥中"K.stuttgartiensis"为主要厌氧氨氧化菌,经过2 a运行该菌在污泥中的丰度由11%提高到24%.     

长期低聚磷条件对AO-SBR系统Accumulibacter代谢特性的影响

为了考察长期低聚磷(Poly-P)条件下Accumulibacter的菌群结构及其代谢特性的变化,采用两个SBR反应器,分别采用乙酸钠和丙酸钠为有机碳源,接种富含Accumulibacter的活性污泥,在进水PO34--P浓度为2. 5 mg·L~(-1)的低磷进水条件下运行60 d,对系统除磷性能、污泥浓度、主要菌群结构的变化进行研究.结果表明,长期低Poly-P条件下两个SBR系统均表现出良好的除磷和去除有机物的性能,但两个系统内的菌群代谢均表现出聚糖菌代谢特征;运行60 d后,丙酸盐系统中AccumulibacterⅠ型仍保持了较高的丰度(40%±7%),表明丙酸盐SBR系统中AccumulibacterⅠ型不仅具有较高的代谢活性,还能够在长期低聚磷条件下以一种不依赖Poly-P的代谢方式进行生存;对比发现,丙酸盐碳源更有利于Accumulibacter适应低磷负荷的运行环境,且在低磷负荷的运行环境下AccumulibacterⅠ型比Ⅱ型更具竞争优势.     

信号分子联合脱氮硫杆菌对同步脱氮除硫的影响

为探究信号分子联合菌对同步脱氮除硫效果的影响,发现同时投加信号分子和脱氮硫杆菌可以加快硫化物和硝酸盐氮的去除且相比单独投加时更有利于单质硫的稳定积累和氮气产量的增加,并通过FISH技术检测了反应结束后微生物总量,信号联合脱氮硫杆菌可以增加微生物总数,因此能够实现较好地脱氮除硫效果.实验从不同信号分子浓度中找出了有利于单质硫稳定积累的最佳浓度,进而在此浓度下分析了单独投加信号分子、单独投加脱氮硫杆菌,以及同时投加信号分子和脱氮硫杆菌3种情况下的脱氮除硫效果.结果表明,当硫化物浓度为200 mg·L-1时,延长反应时间至72 h后,信号分子联合脱氮硫杆菌使硫化物的去除率提高至99. 8%,硝酸盐去除率提高至96. 9%,且单质硫转化为硫酸盐的速率减慢,氮气的产量增加,反应结束后其单质硫和氮气的量分别达到59. 0 mg和80. 0 m L.当硫化物浓度为300 mg·L-1时,单独投加2. 5μmol·L-1的信号分子在72 h时其硫化物和去除率达到99. 0%,硝酸盐的去除达到93. 9%,单质硫和氮气的产量分别达到63. 1 mg和79. 5 m L.     

Prenatal diagnosis of a mosaic supernumerary marker iso (8p) (tetrasomy 8p): discordance between chorionic villi culture and amniotic fluid karyotypes

We describe the first case of mosaic supernumerary marker iso (8p) displaying a karyotype discordance between chorionic villi (CV) and amniotic fluid (AF) cultures during prenatal cytogenetic diagnosis. In the first trimester, cytogenetic analysis after chorionic villi sampling (CVS) was normal in all metaphases in the short-term cytotrophoblast cell culture, but an undefined supernumerary marker was detected in 60% of mesenchymal cells in the long-term CV culture. Informed of the mosaicism, the couple selected amniotic fluid sampling as a second-trimester confirmatory diagnostic procedure. The supernumerary marker was absent in all of the 25 available AF cells metaphases. The prospective parents received genetic counselling and were informed that the discordance could be interpreted as a placental confined mosaicism or as a true foetal mosaicism with low percentage of affected cells. The couple opted to continue the pregnancy. In the second month of life, the child had abnormal development with severe psychomotor delay and frequent episodes of epilepsy. Postnatal cytogenetic extensive re-evaluation discovered that the previously detected supernumerary marker was indeed an isochromosome (8p) rearrangement present at low frequency in 5% of the blood lymphocytes. Copyright © 2006 John Wiley & Sons, Ltd.     

Prenatal and postnatal characterization of Y chromosome structural anomalies by molecular cytogenetic analysis

We describe three cases in which we used fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR) and comparative genomic hybridization (CGH) to characterize Y chromosome structural anomalies, unidentifiable by conventional G-banding. Case 1 was a 46,X,+mar karyotype; FISH analysis revealed an entire marker chromosome highlighted after hybridization with the Y chromosome painting probe. The PCR study showed the presence of Y chromosome markers AMG and SY620 and the absence of SY143, SY254 and SY147. CGH results confirmed the loss of Yq11.2-qter. These results indicated the presence of a deletion: del(Y)(q11.2). Case 2 was a 45,X [14]/46,XY[86] karyotype with a very small Y chromosome. The PCR study showed the presence of Y chromosome markers SY620 and AMG, and the absence of SY143, SY254 and SY147. CGH results showed gain of Yq11.2-pter and loss of Yq11.2-q12. These results show the presence of a Yp isodicentric: idic(Y)(q11.2). Case 3 was a 45,X,inv(9)(p11q12)[30]/46,X,idic(Y)(p11.3?),inv(9)(p11q12)[70] karyotype. The FISH signal covered all the abnormal Y chromosome using a Y chromosome paint. The PCR study showed the presence of Y chromosome markers AMG, SY620, SY143, SY254 and SY147. CGH only showed gain of Yq11.2-qter. These results support the presence of an unbalanced (Y;Y) translocation. Our results show that the combined use of molecular and classical cytogenetic methods in clinical diagnosis may allow a better delineation of the chromosome regions implicated in specific clinical disorders. Copyright © 2002 John Wiley & Sons, Ltd.