亚硝化单胞菌的分离鉴定及其降解特性的研究

对含有亚硝化菌的活性污泥进行富集和分离得到6株亚硝化菌,对其中降解效果最好的一株YS6菌株进行鉴定和降解特性研究。结果表明:该菌为亚硝化单胞菌(Nitrosomonas sp.),最适碳源CaCO_3和NaHCO_3,最适温度30℃,最适pH值8,250mL锥形瓶最佳装瓶量50 mL,接种量12×10~9个。     

两种粒径纳米银对Nitrosomonas europaea的毒性效应

为明晰不同粒径纳米银（AgNPs）对欧洲亚硝化毛杆菌（Nitrosomonas europaea）的毒性效应,采用室内培养方式,探究10nm和50nm的AgNPs对N.europaea生长、氮转化能力、细胞结构、活性氧生成和功能基因表达的影响.结果表明,AgNPs暴露抑制N.europaea生长,随着暴露时间的延长,细菌生长抑制率增加,在4h达到最大值;培养基中NH₄⁺向NO₂^-转化速率减缓,N.europaea的铵态氮转化能力降低;扫描电镜（SEM）图像显示AgNPs造成部分细菌表面塌陷且有孔洞,细胞膜受损严重;透射电镜（TEM）图像显示AgNPs造成细菌内部核物质消融,细胞质膜界限模糊;流式细胞仪（FCM）检测发现AgNPs增加细胞内活性氧的生成;qRT-PCR技术对AgNPs暴露后N.europaea功能基因amoA、hao、merA表达进行测定,发现AgNPs抑制N.europaea功能基因的转录表达.综上所述,AgNPs通过与细胞膜相互作用和产生氧化应激损伤N.europaea,抑制amoA和merA的表达,进而影响铵态氮转化过程,且小粒径AgNPs的毒性强于大粒径.     

Linking NE1545 gene expression with cell volume changes in Nitrosomonas europaea cells exposed to aromatic hydrocarbons

Nitrosomonas europaea, a model ammonia oxidizing bacterium, was exposed to a wide variety of aromatic hydrocarbons in 3 h batch assays. The expression of NE1545, a phenol sentinel gene involved in fatty acid metabolism, was monitored via quantitative real-time polymerase chain reaction (qRT-PCR) and a Coulter Counter technique was used to monitor changes in cell volume. Decreases in cell volume and NE1545 gene expression correlated strongly with exposure to aromatic hydrocarbons that possessed a single polar group substitution (e.g. phenol and aniline). Aromatic hydrocarbons that contain no polar group substitutions (e.g. toluene) or multiple polar group substitutions (e.g. p-hydroquinone) caused negligible changes in NE1545 expression and cell volume. The oxidation of aromatic hydrocarbons by N. europaea from configurations without a single polar group to one with two polar groups (e.g. p-cresol oxidized to 4-hydroxybenzyl alcohol) and from configurations with no polar groups to one with a single polar group (e.g. ethylbenzene oxidized to 4-ethylphenol) greatly influenced NE1545 gene expression and observed changes in cell volume. Nitrification inhibition in N. europaea by the aromatic hydrocarbons was found to be completely reversible; however, the decreases in cell volume were not reversible suggesting a physical change in cell membrane composition. Ammonia monooxygenase blocking studies showed that the chemical exposure that was responsible for the cell volume decrease and up-regulation in gene expression and not the observed inhibition. N. europaea is the first bacterium shown to experience significant changes in cell volume when exposed to μM concentrations of aromatic hydrocarbons, three orders of magnitude lower than previous studies with other bacteria.     

一株高适应性Nitrosomonas eutropha CZ-4的脱氨特性

从垃圾渗滤液中分离得到了一株亚硝化单胞菌Nitrosomonas eutropha CZ-4,其16S rDNA序列与N.eutropha C91的相似性达99%.研究了pH值、温度、游离亚硝酸浓度、盐度等对其生长的影响,并测试了其在垃圾渗滤液、黑臭水和富营养化湖水中的脱氨效果.结果表明,该菌的最适生长pH值为7.3~8.7,最适生长温度为30.9℃,游离亚硝酸和盐度对该菌的半数抑制浓度分别约为0.11mg/L与2%.在最佳发酵条件下,该菌的最大氨氮去除速率为58mg/（L·h）,最短倍增时间为8.2h;在不同类型的污水/地表水（初始氨氮浓度为0.66~603mg/L）中,该菌的最大氨氮去除速率为11.4mg/（L·h）,最短倍增时间为10.9h,最低残余氨氮浓度为0.11mg/L.     

亚硝化细菌的筛选及培养条件的研究

从活性污泥中分离出16株亚硝化细菌,筛选出亚硝酸盐氮积累速率较高的两株菌Y8和Y16,初步鉴定Y8菌株为亚硝化球菌（Nitrosococcns．sp）,Y16菌株为亚硝化单胞菌（Nitrosomonas.sp）,并对其生长曲线进行了测定。通过氨氮去除率或业硝酸盐氮的质量浓度来验证亚硝化细菌的活性,考察了生成的亚硝酸盐氮的质量浓度与培养时间的关系、亚硝化细菌的活性与培养温度的关系、亚硝化细菌的活性与培养基pH的关系。Y8和Y16菌株在30℃、培养基pH为7．5、130r／min的条件下振荡培养5d,氨氮去除率分别为81．12％,75．36％。     

Synthesis of 2-furyl-4-arylidene-5(4H)-oxazolones as new potent antibacterial agents against phyto-pathogenic and nitrifying bacteria

Crop losses due to bacterial pathogens are a major global concern. Most of the available pesticides for these pathogens suffer from various drawbacks such as complicated synthesis, high cost, high toxicity, pesticide resistance and environmental hazards. To overcome these drawbacks, the present study was undertaken to find a potent bactericide. Therefore, a series of compounds comprising bioactive furyl and oxazolone rings was synthesized under microwave irradiation and screened for in vitro antibacterial activity. The reactions were completed in fewer than 2 minutes with minimal use of solvents and resulted in high yields. These compounds were screened for antibacterial activity against plant pathogens, Xanthomonas oryzae, Ralstonia solanacearum and nitrifying bacteria, Nitrosomonas species under laboratory conditions. Five compounds were active as antibacterial agents against Xanthomonas oryzae and Ralstonia solanacearum. However, all compounds were effective against the Nitrosomonas species and the best one was 2-furyl-4-(3-methoxy-4-hydroxybenzylidene)-5(4H)-oxazolone. The study revealed the fast and environmentally friendly synthesis of bioactive title compounds, which also hold promise to be used as prototypes for the discovery of potent analogues.     

Influence of liberated silver from silver nanoparticles on nitrification inhibition of Nitrosomonas europaea

The ecotoxicity of silver nanoparticles (Ag-NPs) to wastewater biota, including ammonia oxidizing bacteria (AOB), is gaining increasing interest as the number of products containing Ag-NPs continues to rise exponentially and they are expected to accumulate in wastewater treatment plants. This research demonstrated that the addition order of Ag-NP and the media constituents had a profound influence on the stability of the Ag-NP suspension and the corresponding repeatability of results and sensitivity of Nitrosomonas europaea. N. europaea, a model AOB, was found to be extremely sensitive to ionic silver (Ag⁺) and two sizes of Ag-NPs (20 and 80 nm). Ag⁺ exposures resulted in the highest level of toxicity with smaller Ag-NPs (20 nm) being more toxic than larger Ag-NPs (80 nm). The increased sensitivity of N. europaea to smaller Ag-NPs was caused by their higher rates of dissolved silver (dAg) release, via dissolution, due to a greater surface area to volume ratio. dAg was shown to be responsible for the vast majority of the observed Ag-NP toxicity, as determined by abiotic Ag-NP dissolution tests. For the sizes of Ag-NP studied (20 and 80 nm), there appears to be a negligible nanoparticle-specific toxicity. This was further supported by similarities in inhibition mechanisms between Ag⁺ and Ag-NP, with both causing decreases in AMO activity and destabilization of the outer-membrane of N. europaea. Finally, equal concentrations of total silver were found to be tightly associated to both Ag⁺ and Ag-NP-exposed cells despite Ag-NP concentrations being five times greater, by mass, than Ag⁺ concentrations.     

Effects of selected pharmaceutically active compounds on the ammonia oxidizing bacterium Nitrosomonas europaea

Pharmaceutically active compounds (PhACs) are commonly found in wastewater influent. However, little research has focused on determining their impact on fundamental processes in wastewater treatment such as nitrogen removal. In this study, focus was placed on 4 commonly occurring PhACs (ketoprofen, naproxen, carbamazepine and gemfibrozil). Their effect was ascertained in the ammonia oxidizing bacterium (AOB), Nitrosomonaseuropaea in terms of membrane integrity and nitrite production. These PhACs were shown to inhibit nitrite production at concentrations of 1 and 10 μM while no effect was observed at 0.1 μM. The maximum observed nitrification inhibition was 25%, 29%, 22% and 26% for ketoprofen, naproxen, carbamazepine and gemfibrozil, respectively. A decrease in the live/dead ratio ranging from 10% to 16% suggests that these PhACs affect membrane integrity in N.europaea. The difference in nitrite production between PhACs treated cells and non PhAC treated controls was still significant following washing suggesting that inhibition is irreversible. Finally, nitrite production when adjusted to the live fraction of cells was also found to decrease suggesting that PhACs inhibited the activity of surviving cells. These results suggest that the presence of PhACs may affect AOB activity and may impact nitrogen removal, a key function in wastewater treatment. Follow up studies with additional AOB and in mixed culture are needed to further confirm these results.