饮用水微生物的安全快速检测

【目的】为了更好地分析饮用水中的微生物含量。【方法】利用流式细胞术(Flowcytometry,FCM)、ATP测定方法检测瓶装无气饮用水中的微生物数量、可同化有机碳(Assimilable organic carbon,AOC)含量以及微生物活性,并将检测结果与传统的饮用水微生物检测技术相对照。【结果】FCM方法可快速区分水样中的活性细菌和非活性细菌,AOC含量反映了水样中微生物再生能力;而ATP检测方法也能比异养细菌平板计数法(Heterotrophic plate count,HPC)更好地反映瓶装无气饮用水中的实际微生物含量。【结论】FCM、ATP测定方法要明显优于依赖于培养的传统方法。     

Molecular Identification and Biofilm-Forming Ability of Culturable Aquatic Bacteria In Microbial Biofilms Formed in Drinking Water Distribution Networks

Drinking water distribution networks are known to harbor microbial biofilms. The aim of the present work is to (i) identify the culturable bacteria presented in the drinking-water distribution network, (ii) investigate the ability of isolated bacteria to form biofilm under some environmental stress conditions and some eliminating or removing treatments. To achieve it, 57 strains were isolated from biofilm (43 isolates) and water samples (14 isolates) collected from five stations in drinking-water distribution network in Taif city, Kingdom of Saudi Arabia (KSA). Partial sequences of 16S rRNA gene in the 57 isolates ensured the presence of only 22 different strains in biofilm samples. Among these strains, only 14 strains were also detected in water samples. Gram-negative Aeromonas hydrophila was the most occurred bacterium in the microbial biofilm obtained from the purified-water storage tanks followed by Gram-negative Pseudomonas sp. Gram-positive Bacillus subtilis was the most occurred bacterium in the microbial biofilm collected from the ends of the distribution pipes. Among the 22 isolated strains, 13 strains were strong biofilm producers at 30 and 37°C. The effects of environmental stresses including nutrient starvation (diluted TSB, 20:1), heating (100°C for 10 min), UV-treatment (240 nm for 10 min) and dynamic incubation (150 rpm min^?1) on the formation of biofilm were also investigated. These conditions affected the biofilm formation ability of the isolated strains at different levels. Nutrient starvation enhanced biofilm formation by most of the isolates. Among some biofilm deforming treatments, SDS and trypsin had considerable effects on preventing biofilm formation by most of the isolated strains. In conclusion, the results of the present work indicated that not all biofilm strains released from biofilm to the drinking water. Also, not all biofilm strains were able to form biofilm. Most of isolated bacteria had ability to form biofilm at suboptimum temperature of growth. These results may provide basic information on formation of microbial biofilms and overcome the problem of deteriorating of water quality in the drinking-water distribution networks.     

醋酸菌多相分类研究进展

醋酸菌是一大群革兰氏染色阴性、绝对好氧的细菌的总称, 能将乙醇或糖类不完全氧化为有机酸。醋酸菌的分类在近30年经历了很大变化, 早期的分类系统主要以表型和生化特征为基础。如今, 大多采用结合表型、化学分类法和基因型数据的多相分类法对醋酸菌进行分类。本文综述了醋酸菌的多相分类研究进展, 主要介绍了醋酸菌的现行分类情况及表型分类、化学分类和基因分型等方法在醋酸菌分类中的应用。     

基因工程菌培养过程中乙酸的积累及其对工程菌生长和产物表达的影响

在rIL-2工程菌K802(pLY-4)高密度培养中发现培养液中有大量代谢副产物乙酸积累,乙酸的存在对工程菌的生长和产物的表达均有明显的抑制作用,这种抑制作用是制约工程菌高密度培养的重要因素,为了减小这种抑制作用,研究了培养基pH与乙酸抑制作用的关系,发现适当提高培养基pH值,能有效地减小乙酸的抑制作用;高密度培养时,提高培养基的pH后,虽然仍有大量乙酸积累,但产物的表达水平和菌密度都有一定的提高     

马尾松人工林土壤各粒径团聚体湿筛后的有机碳分配

选取25a、45a和65a马尾松人工林为研究对象,采用湿筛法对各粒径土壤团聚体分别湿筛。探究了马尾松人工林各粒径团聚体湿筛后的团聚体有机碳分配,以探讨各粒径团聚体湿筛后分配到同一粒级团聚体有机碳含量及其对团聚体水稳性的贡献差异。结果表明：种植年限增加显著降低土壤团聚体水稳性（P<0.05）;各粒径团聚体湿筛后分配的有机碳随粒级减小含量呈先降后增趋势,以保持原粒级团聚体有机碳（12.96-32.01 g/kg）含量最高,其次是<0.25 mm粒级（8.08-23.53 g/kg）。各粒径团聚体湿筛分配到同一粒级的有机碳以保持原粒级的含量最高（P<0.05）;土壤团聚体水稳性与各粒径团聚体湿筛后保持原粒径的有机碳呈显著或极显著正相关（P<0.05或0.01）,分配到越小的粒级正相关性越不显著。此外,团聚体水稳性与各粒径团聚体湿筛分配到同一粒级的有机碳呈正相关,以保持原粒级相关性最高（P<0.01或0.05）;回归方程及相关性系数表明,有机碳与保持原粒径团聚体呈显著呈或极显著正相关（P<0.05或0.01）,与消散到其他粒级的团聚体呈负相关或极显著负相关（P>0.05或<0.01）。本研究得出有机碳含量增加促进更大粒径团聚体形成。反之,促使大粒径团聚体向较小粒径团聚体转化。同一粒级团聚体间,保持原粒级团聚体比易转化形成更大粒级团聚体有更高的有机碳含量和更强的水稳性,这对团聚体的固碳提供了新的认识。     

Bacterial species in biofilm cultivated from the end of the Seoul water distribution system

AIMS: To investigate changes in the bacterial population and the safety of the biofilm at the end of the drinking water distribution system in Seoul (Korea), selective media and bacterial community analyses were applied to a semi-pilot galvanized iron pipe (GIP) model. METHODS AND RESULTS: No total coliforms or faecal streptococci were detected on m-Endo or m-Enterococcus agar. No Salmonella spp. and Shigella spp. were detected on bismuth sulphite agar or Hektoen enteric agar, respectively. The latter two media detected coliforms, where m-Endo was negative. Biofilm formation started within 1 week (ca 104 CFU cm(-2)) and exceeded 105 CFU cm(-2) within 6 weeks. Although the fatty acid methyl ester analysis revealed dynamic changes in bacterial composition, Micrococcus, Bacillus, and Pseudomonas spp. were persistent members of the biofilm community. Micrococcus spp. was detected most frequently and in high numbers. CONCLUSIONS: Coliforms and Enterococcus species can be recovered from biofilms in water distribution systems. SIGNIFICANCE AND IMPACT OF THE STUDY: This study illustrates the role of biofilms in the chronic deterioration of the water-distribution system in Seoul (Korea).     

生态恢复对红壤侵蚀地土壤有机碳组成及稳定性的影响

为了研究红壤侵蚀区生态恢复过程中土壤有机碳的组成与动态变化,选择红壤侵蚀区生态恢复10 a和30 a的马尾松林为对象,以侵蚀裸地和次生林为对照,应用土壤有机碳物理分组方法,研究了侵蚀地植被恢复过程中表层土壤粗颗粒态有机碳(cPOC)、细颗粒态有机碳(f POC)和矿质结合有机碳(MOC)含量及POC/MOC比值的变化。结果表明:生态恢复显著提高了土壤有机碳含量(P0.05),土壤中不同组分有机碳含量也相应增加。生态恢复10 a,土壤有机碳主要以f POC形式积累,cPOC和MOC没有显著变化,其中0—10 cm土层POC占总土壤有机碳(SOC)比例高达64.1%,但稳定性较差。与恢复10 a相比,生态恢复至30 a时,0—10 cm土壤f POC含量相对不变,cPOC和MOC含量均显著增加(P0.05),10—20 cm土壤f POC和MOC增加量达到显著水平,而cPOC含量仍未显著增加,说明生态恢复过程中土壤固碳模式符合SOC饱和理论。生态恢复过程中土壤POC/MOC比值呈先升高后降低的趋势,且表层土壤大于亚表层土壤,说明随着生态恢复时间的增加,土壤有机碳稳定性逐渐提高,且亚表层土壤高于表层。因此,生态恢复对于侵蚀地碳固定的长期有效性具有重要意义。     

The effect of material choice on biofilm formation in a model warm water distribution system

Water distribution systems (WDS) are composed of a variety of materials and may harbour potential pathogens within surface-attached microbial biofilms. Biofilm formation on four plumbing materials, viz. copper, stainless steel 316 (SS316), ethylene propylene diene monomer (EPDM) and cross-linked polyethylene (PEX), was investigated using scanning electron microscope (SEM)/confocal microscopy, ATP-/culture-based analysis, and molecular analysis. Material ‘inserts’ were incorporated into a mains water fed, model WDS. All materials supported biofilm growth to various degrees. After 84 days, copper and SS316 showed no significant overall differences in terms of the level of biofilm formation observed, whilst PEX supported a significantly higher level ofbiofilm. EPDM exhibited gross contamination by a complex, multispecies biofilm, at a level significantly higherthan was observed on the other materials, regardless of the analytical method used. PCR-DGGE analysis showed clear differences in the composition of the biofilm community on all materials after 84 days. The primary conclusion of this study has been to identify EPDM as a potentially unsuitable material for use as a major component in WDS.     

Effects of biological soil crusts on profile distribution of soil water,organic carbon and total nitrogen in Mu Us Sandland,China

Aims Biological soil crusts (BSCs) can affect soil properties including water dynamics and cycling of soil carbon and nitrogen in dryland ecosystems. Previous research has mostly focused on effects of BSCs on soil water distribution or carbon and nitrogen fixation in the surface soil layer. Thus, little is known about effects of BSCs on properties throughout the soil profile. In the current study, we assessed the effects of BSCs on the distribution of soil water content (SW), soil organic carbon content (SOC) and soil total nitrogen content (STN) throughout the soil profile as well as the influence of water conditions on the effects of BSCs.Methods In a field investigation in Mu Us Sandland, North China, soil samples were taken from plots with and without BSCs on 13 and 28 September 2006, respectively. On the two sampling dates, average soil gravimetric water content was 3.83% (±1.29%) and 5.08% (±0.89%), respectively, which were regarded as low and high water conditions. Soil samples were collected every 5 cm to a depth of 60 cm, and SW, SOC and STN were measured in the laboratory.Important findings (i) BSCs affected profile distribution of SW, SOC and STN. In addition, water conditions within the plots significantly modified BSCs' effects on the profile distribution of SW, but marginally affected the effects on SOC and STN. (ii) Under high water conditions, SW in the surface soil layer (0–10 cm) was higher in soils with BSCs compared to those without BSCs, while the opposite was true in the deep soil layer (30–55 cm). (iii) Under low water conditions, SW was lower with BSCs compared with no BSCs in near-surface (5–20 cm) and deep (25–40 cm) soil layers. (iv) BSCs affected SOC and STN only in the surface soil layer (0–5 cm) and were modified by plot water conditions.     

水位和施氮对若尔盖高寒湿地土壤非共生固氮的影响

高寒湿地中土壤微生物固氮是氮元素进入生态系统的主要途径之一,环境因子变化对土壤固氮功能的影响仍不明晰。在四川若尔盖高寒湿地搭建了由27个生态模拟箱组成的中宇宙实验系统,通过控制水位和模拟氮沉降,研究水位变化和施氮对土壤非共生固氮的影响。实验设计了3水位水平×3施氮水平共9个处理,测定了生态模拟箱中表层土壤的非共生固氮速率,土壤碳、氮含量,以及地上植物生物量和植物氮含量,比较不同水位和施氮处理下非共生固氮速率的变化规律并分析其与土壤和植物碳、氮含量的关系。研究发现：生态模拟箱中土壤非共生固氮速率范围是0.003-7.35 μg N g^-1 d^-1,从不淹水到淹水的处理土壤非共生固氮速率提高约2倍。施氮处理中固氮速率随土壤含水量升高而增强的敏感性高于施氮对照处理,且施氮处理下的生态模拟箱中土壤有机碳含量显著升高,据此推测施氮可能使淹水的生态模拟箱中的浮游植物提高生产力而释放可利用有机碳,从而间接促进土壤非共生固氮。本研究获得以下结论：（1）若尔盖高寒湿地中土壤水位是限制固氮速率的重要因子;（2）施氮背景下土壤含水量对非共生固氮的促进效应更明显。     

高温胁迫对~(14)C-水杨酸在葡萄苗中运转分配的影响

用 38℃高温和 2 7℃正常温度分别处理14 C SA标记的葡萄幼苗叶片 1,3,6 ,12h ,发现常温下14 C SA向外运转的很少 ,而高温下向外运转的更少。高温处理和常温对照的14 C SA都既向上运输又向下运输 ,以向下运输为主 ,但无论向上运输还是向下运输 ,高温下要比常温下运输的少一些。在常温下 1,3,6h ,14 C SA以根系分配的最多 ,12h以茎中分配最多 ,而在高温处理下 1,3h以根中分配最多 ,6 ,12h则以茎中分配最多。高温处理和常温对照下都是离引入叶越近的茎段和叶片 (除引入叶 ) ,14 C SA含量越高     

大颗粒活化腐植酸肥对苹果土壤团聚体和有机碳的影响

新型大颗粒活化腐植酸肥(LAF)在苹果化肥减量和果实稳产方面的效果显著,探明其对土壤团聚体和有机碳的影响是揭示该新型肥料对苹果土壤结构影响的重要依据。本研究设置4个LAF处理:LAF₁[全量施肥,施肥时期及重量比(下同):萌芽期∶膨果期∶成熟期=3∶4∶3]、LAF₂(全量施肥,萌芽期∶膨果期∶成熟期=2∶3∶5)、LAF₃(减量1/4施肥,萌芽期∶膨果期∶成熟期=2∶3∶5)、LAF₄(减量1/3施肥,萌芽期∶膨果期∶成熟期=2∶3∶5),以不施肥(CK)处理为对照。通过4年盆栽试验,研究苹果土壤团聚体组成、稳定性和有机碳对不同施肥处理的响应。结果表明: 1)与CK相比,LAF各处理显著提高了土壤水稳性大团聚体含量,>2 mm和2~0.25 mm粒径团聚体含量分别提高了53.4%~77.5%和12.3%~17.0%,且提高幅度随施肥量的增加而增大,其中LAF₁处理土壤水稳性大团聚体含量最高。2)LAF各处理在各粒径团聚体含量上差异不显著,其中2~0.25 mm粒径团聚体含量所占的比例最高。3)与CK相比,LAF各处理均显著提高了团聚体平均重量直径(MWD)和几何平均直径(GMD),降低了分形维数(D),其中LAF₁处理的MWD和GMD值最高,对土壤团聚体稳定性提升效果最好。4)除LAF₄外,其他LAF处理土壤有机碳含量均显著高于CK,其中LAF₂处理土壤有机碳含量最高;LAF各处理均增加了土壤各粒径团聚体有机碳含量,LAF₁、LAF₂、LAF₃处理显著提高了>2 mm粒径团聚体有机碳含量,且>2 mm团聚体有机碳对总有机碳的贡献率最大;LAF各处理的水稳性大团聚体有机碳对总有机碳的贡献率均显著高于CK,且贡献率均在66.0%以上,其中LAF₁处理最高。综上,施用LAF在促进苹果土壤水稳性大团聚体形成和稳定性、提高团聚体有机碳含量上应用效果显著,其中全量施用效果最好。施用LAF可作为改善苹果土壤结构和提升土壤肥力的有效措施。     

自然风化过程对赤泥团聚体有机碳组分的影响

以华中地区某氧化铝企业赤泥堆场为研究对象,采集不同时间赤泥堆场(1a,10a,20a)0—20 cm样品,采用物理分组方法,研究自然风化过程对赤泥团聚体有机碳组分的影响。结果发现,赤泥轻组有机碳分配比例随堆存时间的延长呈上升趋势。重组有机碳约占赤泥总有机碳的97.24%—99.11%,以粗颗粒有机碳和矿物结合态颗粒有机碳为主。3种堆存时间(1a,10a,20a)赤泥团聚体颗粒有机碳含量变化范围分别为1.21—1.85、2.62—2.95、3.52—4.15 g/kg。颗粒有机碳含量在2—1 mm粒级赤泥团聚体中最高,随赤泥粒级减小而降低,其中矿物结合态颗粒有机碳分配比例最高,游离态颗粒有机碳分配比例最低。这表明自然风化过程增加了赤泥有机碳组分含量,提高了赤泥中有机碳库的稳定性。     

增氮对青藏高原东缘典型高寒草甸土壤有机碳组成的影响

土壤有机碳动态是陆地生态系统碳平衡研究的关键环节,有关青藏高原高寒草甸土壤有机碳组成对大气氮沉降增加的响应研究至今尚未开展。基于中国科学院海北生态站的大气氮沉降模拟控制实验平台,于2010年5月、7月和9月中旬分别测定不同施氮处理下0—10cm、10—20cm、20—30cm土壤中粗颗粒态有机碳(CPOC)、细颗粒态有机碳(FPOC)和矿质结合有机碳(MOC)含量,研究不同施氮类型(NH4Cl,(NH4)2SO4和KNO3)和施氮水平(0、10、20、40 kgN.hm-.2a-1)对土壤POC和MOC含量以及POC/MOC比值的影响。结果表明:青藏高原高寒草甸土壤POC积聚在土壤表层,占总土壤有机碳(SOC)含量的64%以上,稳定性较差。施氮水平显著改变了土壤CPOC、FPOC和MOC含量,而施氮类型的影响不显著。不同月份土壤POC和MOC含量对增氮的响应不同,反映了SOC组分对增氮响应的时间异质性。在生长季中期,施氮倾向于增加表层土壤POC含量,而在生长季初期和末期恰好相反。土壤MOC对增氮的响应不敏感。另外,施氮显著降低生长季初期表层土壤POC/MOC比例,SOC稳定性增加。表明,青藏高原高寒草甸土壤有机碳活性组分较高,未来大气氮沉降增加短期内即可降低活性有机碳含量,相应地改变了其组成和稳定性。     

生物光子场的光谱分布特性研究

改装设计了分光谱段测试生物光子场辐射强度的装置;用此装置实验证明生物系统的超微弱光子辐射（ＵＰＥ）存在于从紫外－可见到红外的宽光谱范围内,而且其ｆ（λ）≌常数。此结果对于证实生物光子场的相干特性具有一定意义。     

不同稻作制对红壤性水稻土中锰剖面分布的影响

为了探讨红壤性水稻土锰的迁移和转化行为,通过长达16a的定位试验研究了不同稻作制、有机肥以及地下水位对土壤剖面中全锰、活性锰和交换态锰分布的影响。试验结果表明,长期淹水种稻引起0-20cm土壤层次全锰、活性锰和交换态锰含量的显著下降,而在20－40cm和40－75cm土层相对累积。不同稻作制比较,0－20cm土层中全锰、活性锰和交换态锰含量以稻稻泡显著高于稻稻绿和稻稻油处理,而20－40cm和40－75cm土层3种锰形态的含量各稻作制之间无显著差异,表明实行水旱轮作的稻稻绿与稻稻油两种稻作制耕层土壤锰的淋溶损失比持续淹水的稻稻泡制更为严重。相对而言,不同有机肥施用水平和地下水位对土壤剖面中锰分布的影响要小于稻作制,总的趋势为：土壤剖面中锰的空间分异程度以高量有机肥＞常量有机肥＞单施化肥;低水位＞高水位。从土壤中锰的空间分布规律可以看出,水旱轮作（尤其是在低水位和有机无机配合的条件下）比长期淹水更有利于土壤锰氧化还原引起的深层淋溶与淀积,加速了典型水稻土剖面的形成。     

外源ABA和水分胁迫对不同基因型三叶草生长的影响

在室内研究了不同浓度外源ABA处理和不同水分胁迫对6种不同基因型的三叶草(Trifolium subterraneum L.)生长的影响.当三叶草的第四片叶完全展开时,向营养液中施加不同浓度的ABA时对盆栽土壤进行控水.在处理的1,4,7和11 d,测定植株鲜重、叶片数、最长根长,以表示三叶草的生长状况.各参试品种以上三项生长指标均受外源ABA和水分胁迫的影响而呈现下降的趋势.同时,叶片水势值随ABA浓度的增加和水分胁迫强度的增加而明显降低.在10-4 mol/L ABA处理11 d后,参试品种平均生长量的减少与水分胁迫15 d后其生长量的减少的结论一致.在不同浓度ABA处理下,不同基因型三叶草平均叶片数,完全展开叶面积和每株干物质重约降低了50%,而其根冠比却增加了80%.不同基因型三叶草生长参数间的变化及排序结果与盆栽相同品种获得的实验结果非常相似.品种Clare、Nuba 和Seaton Park在对照和处理下均表现最好.由于三叶草对一定浓度范围的外源ABA的反应与其在盆栽水分胁迫下的反应结果十分相似,因此,利用外源ABA处理的方法来研究不同基因型三叶草的耐旱性将可能是一种行之有效的方法.     

外源ABA和水分胁迫对不同基因型三叶草生长的影响

在室内研究了不同浓度外源ABA处理和不同水分胁迫对 6种不同基因型的三叶草 (TrifoliumsubterraneumL .)生长的影响。当三叶草的第四片叶完全展开时 ,向营养液中施加不同浓度的ABA时对盆栽土壤进行控水。在处理的 1,4 ,7和 11d ,测定植株鲜重、叶片数、最长根长 ,以表示三叶草的生长状况。各参试品种以上三项生长指标均受外源ABA和水分胁迫的影响而呈现下降的趋势。同时 ,叶片水势值随ABA浓度的增加和水分胁迫强度的增加而明显降低。在 10 -4mol/LABA处理 11d后 ,参试品种平均生长量的减少与水分胁迫 15d后其生长量的减少的结论一致。在不同浓度ABA处理下 ,不同基因型三叶草平均叶片数 ,完全展开叶面积和每株干物质重约降低了5 0 % ,而其根冠比却增加了 80 %。不同基因型三叶草生长参数间的变化及排序结果与盆栽相同品种获得的实验结果非常相似。品种Clare、Nuba和SeatonPark在对照和处理下均表现最好。由于三叶草对一定浓度范围的外源ABA的反应与其在盆栽水分胁迫下的反应结果十分相似 ,因此 ,利用外源ABA处理的方法来研究不同基因型三叶草的耐旱性将可能是一种行之有效的方法。