Assessment of metabolic activity of single bacterial cells — new developments in microcolony and dehydrogenase assays

Abstract: Some bacteria lose culturability in natural environments but retain measurable metabolic activity and are thus considered viable. Several techniques have been proposed to determine the activity of nonculturable cells. Due to the considerable physiological heterogeneity of bacterial populations in the environment, it is imperative to apply methods which measure cellular activity at the single cell level. This review focuses on two promising methods: the microcolony assay and the respiration assay based on reduction of 5-cyano-2,3-ditolyl tetrazolium chloride (CTC). In the microcolony assay, viable cells are identified by their ability to perform a limited number of cell divisions and this approach is thus related to conventional culture techniques. Some recent methodological developments of the technique aiming at improving the incubation conditions and the detection of microcolonies are presented. Results obtained by the microcolony technique are used to introduce its advantages and limitations. The CTC-reduction assay determines a central cellular metabolic activity, but does not measure cell growth. Results of studies using this assay are presented, and it is emphasized that great care should be taken to optimize assay conditions for the studied organisms. Finally, the results obtained by different viability assays are compared. For a specific bacterium, several assays, addressing different aspects of cell metabolism, can provide comparable results suggesting that they provide meaningful viability estimates. On the other hand, the use of viability assays on complex indigenous populations may be ambiguous.     

Hydraulic lift of Medicago sativa and Astragalus laxmannii and its effect on their neighborhood plants

为探讨紫花苜蓿(Medicago sativa)及斜茎黄耆(沙打旺, Astragalus laxmannii)与禾本科牧草混播后的水力提升现象, 揭示深、浅根性牧草的种间关系, 为混播草地的建植提供理论依据, 该研究开展了室外“上下盆”分根盆栽试验、采用土壤水分测定及“重水” (D₂O, 氘(D)含量>99.9%)标记法估算了苜蓿及斜茎黄耆分别与‘冬牧70’黑麦(Secale cereal ‘Dongmu 70’)不同混播比例(豆科:禾本科分别为3:7、5:5、7:3)条件下水力提升的发生情况及其对伴生牧草生长生理性状的影响。结果表明: ‘冬牧70’黑麦与斜茎黄耆混播后的产量显著高于其与紫花苜蓿混播后的产量, 同一种禾豆牧草混播组合不同混播比例中, 以AC2 (紫花苜蓿:‘冬牧70’黑麦为5:5)和BC3 (斜茎黄耆:‘冬牧70’黑麦为7:3)混播组合的总产量最高。不同单混播组合的单株整个生育期内日均提水量存在显著差异, 两种豆科牧草在混播时日均提水量均高于单播时, 斜茎黄耆单混播时的日均提水量显著高于紫花苜蓿, BC2组合(斜茎黄耆:‘冬牧70’黑麦为5:5)的日均提水量高于其他混播组合。在用标记水处理下盆土壤后, 各组合上下盆土壤水氢稳定同位素比率(δD)值显著升高。不同禾豆牧草组合上盆土壤水δD及禾本科牧草茎秆水δD、整株碳同位素分辨率(Δ¹³C)和产量数据表明, 在斜茎黄耆与‘冬牧70’黑麦混播比例为3:7、紫花苜蓿与‘冬牧70’黑麦混播比例为5:5时, 禾本科牧草水分状况或产量好于其他混播比例。以上结果表明, 两种深浅根豆科牧草与浅根性禾本科牧草混播种植时发生了水力提升现象, 两种豆科牧草提升的水分可以被伴生的禾本科牧草所吸收利用。     

Effects of Inorganic Oxides,Sulfides, Chlorides,and Acid Chlorides on Metachromasia and Other Staining Properties

The ability of 17 inorganic compounds (POCl₃, PSC1₃, PC1₃, P₂O₅, P₂S₅, P₄S₃, P₄S₇, PC1₅, Sb₂O₅, As₂O₅, BiOC1₂, SeOC1₂, SO₂C1₂, Sb₂S₅, VOC1₂, SiC1₄ and CrO₂Cl₂) dissolved in pyridine or 2,2,4-trimethyl pentane, to enhance subsequent staining of tissue components with toluidine blue, phosphotungstic acid-hematoxylin (PTAH), leukofuchsin, and dihydroxydinaphthyl-disulfide (DDD) was studied. Eight of these compounds were also tested for ability to enhance staining with Alcian blue 8GN and Luxol fast blue MBS. Nine of the 17 compounds produced increased staining of certain tissue components with leukofuchsin, 13 with toluidine blue, 16 with PTAH, and 16 with DDD. The results suggest additional approaches to identification of tissue entities by induced metachromatic basophilia and leukofuchsin positivity as well as by the other stains studied, and also suggest a number of hitherto unstudied modes of reaction between the dyes used and reactive groups of tissue components. Many reactions of the compounds tested, with reactive groups known to be present in tissue components, are basecatalyzed, so that choice of solvent can influence the results obtained.     

Novel complex hydrogels based on N-carboxyethyl chitosan and quaternized chitosan and their controlled in vitro protein release property

A novel pH-responsive hydrogel (CHC) composed of N-carboxyethyl chitosan (CEC) and N-[(2-hydroxy-3-trimethylammonium) propyl] chitosan chloride (HTCC) was synthesized by the redox polymerization technique. Turbidimetric titrations were used to determine the stoichiometric ratio of these two chitosan derivatives. The hydrogel was characterized by FT-IR, thermal gravimetric analysis (TGA), X-ray diffractometry (XRD), and scanning electron microscopy (SEM). The dynamic transport of water showed that the hydrogel reached equilibrium within 48 h. The swelling ratio of CHC hydrogel depended significantly on the pH of the buffer solution. The performance of the CHC as a matrix for the controlled release of BSA was investigated. It was found that the release behavior was determined by pH value of the medium as well as the intermolecular interaction between BSA and the hydrogels.     

环境介质中铂族金属形态分析方法的研究进展

环境中铂族金属(PGMs)的赋存形态多样,形态分析对识别其生态风险具有十分重要的意义。本文综述了环境中3种主要铂族金属(铂、钯、铑)的形态分析方法,包括化学顺序提取、仪器联用技术及计算机模拟等,概述了这些方法的类型、特点及应用,同时阐述了它们存在的不足,并对未来发展方向进行了展望。化学顺序提取法普遍用于固相样品形态分析,当前研究中提出的提取条件和步骤多样,但不能很好地标准化;仪器联用技术在溶液元素形态分析上具有显著优势,毛细管电泳联用系统能够分离具有相同电泳能力的相似物质,但在分离能力和检出限方面不如液相色谱联用系统;计算机模拟则进一步拓展了形态分析的途径,能够实现复杂的形态计算。建议今后将多个方法进行结合,通过相互补充与完善,不断提高分析技术准确性。     

Dispersal of mass‐reared sterile,laboratory‐domesticated and wild male Queensland fruit flies

Queensland fruit flies, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) (‘Q‐flies’) were released as sexually immature adults from a point within an orchard. Marked male Q‐flies were recaptured in the trap furthest from the release point (1087 m) by 2 weeks after release, although 98.25 ± 1.04% of recaptured males were trapped <500 m from the release point. Comparison of gamma‐irradiated (sterile), laboratory‐adapted and wild male Q‐flies indicated that dispersal distance was not significantly affected by fly type. There was no significant correlation between temperature and mean dispersal distance, but total recaptures were significantly negatively correlated with increasing daily maximum, minimum and average temperature.     

微卫星技术分析地方鸡种群体的遗传结构

利用微卫星技术和14对引物MCW150、MCW134、MCW145、MCW104、ADL210、MCW88、ADL237、ADL201、ADL289、MCW5、MCW217、MCW29、ADL176、MCW4对7个地方鸡种和1个引进鸡种的群体遗传结构特征进行了分析。根据池DNA所获得的品种间相似指数计算遗传距离。结果表明:AA鸡与其它品种鸡遗传距离最远,白耳鸡次之,其余品种间较近。这些鸡种的遗传结构特征与其地理分布有关,说明微卫星技术作为畜禽遗传结构分析的辅助手段是可行的,也是高效的 。     

低能离子束在生物技术中的应用研究

自从发现离子注入生物效应后,低能离子与生物体系相互作用研究在我国率先兴起,并很快投入应用。简要介绍低能离子注入生物效应的机理研究和应用研究的进展状况,并展望未来 。     

基于变分方法的内蒙古典型草原水热通量估算

利用内蒙古典型草原生长季(2004年8月13～18日、2005年5月21～26日)和非生长季(2004年12月10～15日)共18 d的涡动相关系统观测资料(10 Hz输出1次)和小气候梯度系统在线输出资料(0.5 h输出1次),分析了变分方法对草原陆气通量估算的准确性.结果表明,变分法估算的感热与潜热通量与涡动相关法观测结果的变化趋势较为一致,且能量闭合程度更高.变分法计算的感热通量在白天明显地较涡动相关法得到的通量值高,12:00前后两者通量都达到峰值,两者之差也达到峰值,而夜间则较为接近;变分法计算的潜热通量曲线的相位略微落后于涡动相关法的通量曲线,且夜间的涡动相关法测得的潜热通量负值极少,负值通量的绝对值也很小.这表明夜间大气稳定导致涡动相关法存在一定程度的通量低估现象.变分法与波文比能量平衡法对陆气通量的估算比较表明,变分法可避免能量平衡法计算不稳定导致的虚假峰值现象,计算结果较为稳定.     

The value of direct fluorescent antibody (DFA) testing for the detection of Pneumocystis Carinii In cytological specimens

A direct fluorescent antibody (DFA) method was compared with methenamine silver staining (MSS) for the detection of Pneumocystis carinii in 384 cytological specimens. DFA testing was more sensitive than the MSS, with P. carinii detected in 31 specimens with DFA and 24 with the MSS. Results of the two methods disagreed in 17 specimens, all of which were sputa. Twelve sputum specimens were DFA positive/MSS negative and five were MSS positive/DFA negative. It is concluded that the DFA technique, although relatively expensive, is simple to perform and offers superior sensitivity to the MSS. However, in sputum specimens the combined use of DFA and MSS leads to optimal sensitivity for the detection of P. carinii.