Rapid detection of Escherichia coli in waters using fluorescent in situ hybridization,direct viable counting and solid phase cytometry

Aims: We developed an improved Fluorescent In Situ Hybridization FISH‐based method to detect viable Escherichia coli cells by solid phase cytometry (SPC), and results were compared to those obtained by the standard culture method. Methods and Results: The method includes a direct viable count (DVC) assay, multi‐probes labelled and unlabelled (helpers) to detect specifically viable E. coli cells and to enhance SPC cell counts. We demonstrate that helpers increase the fluorescence intensity of hybridized E. coli cells as detected by SPC and assess the high specificity of the DVC–FISH procedure on a large panel of cultured strains. Application to seawater, freshwater and wastewater samples showed a good correlation between SPC cells counts and standard plate counts. Conclusion: The high specificity of the procedure was demonstrated as well as its accuracy for detecting and counting viable E. coli cells in environmental samples. Significance and Impact of the Study: The developed approach may be used to monitor faecal contamination sources and to investigate the occurrence of viable E. coli in natural environments.     

生态环境需水量研究进展与理论探析

通过对国内外生态环境需水量研究现状的分析 ,阐述了生态环境需水量的概念。提出生态环境需水量是由河道内生态环境需水和河道外生态环境需水、景观娱乐需水三部分组成的观点 ,从生态环境需水的时空分布、自然生态优先和可持续角度对其内涵进行了探讨 ,分析了生态环境需水量的动态性、可控制性和极限特性。对目前采取的各种生态环境需水量计算评价方法进行了总结 ,展望了该研究领域的发展趋势。     

Formation of nonculturable Escherichia coli in drinking water

AIMS: To examine whether incubation of Escherichia coli in nondisinfected drinking water result in development of cells that are not detectable using standard procedures but maintain a potential for metabolic activity and cell division. METHODS AND RESULTS: Survival and detectability of four different E. coli strains were studied using drinking water microcosms and samples from contaminated drinking water wells. Recovery of E. coli was compared using different cultivation-dependent methods, fluorescence in situ hybridization (FISH) using specific oligonucleotide probes, direct viable counts (DVC), and by enumeration of gfp-tagged E. coli (green fluorescent protein, GFP). Two levels of stress responses were observed after incubation of E. coli in nondisinfected drinking water: (i) the presence of cells that were not detected using standard cultivation methods but could be cultivated after gentle resuscitation on nonselective nutrient-rich media, and (ii) the presence of cells that responded to nutrient addition but could only be detected by cultivation-independent methods (DVC, FISH and GFP). Collectively, the experiments demonstrated that incubation for 20-60 days in nondisinfected drinking water resulted in detection of only 0.7-5% of the initial E. coli population using standard cultivation methods, whereas 1-20% could be resuscitated to a culturable state, and 17-49% could be clearly detected using cultivation-independent methods. CONCLUSIONS: Resuscitation of stressed E. coli on nonselective nutrient-rich media increased cell counts in drinking water using both traditional (CFU), and cultivation-independent methods (DVC, FISH and GFP). The cultivation-independent methods resulted in detection of 10-20 times more E. coli than the traditional methods. The results indicate that a subpopulation of substrate-responsive but apparent nonculturable E. coli may develop in drinking water during long-term starvation survival. SIGNIFICANCE AND IMPACT OF THE STUDY: The existence of substrate-responsive but nonculturable cells should be considered when evaluating the survival potential of E. coli in nondisinfected drinking water.     

The evolution of sex differences in mate searching when females benefit: new theory and a comparative test

Sexual selection is thought to have led to searching as a profitable, but risky way of males obtaining mates. While there is great variation in which sex searches, previous theory has not considered search evolution when both males and females benefit from multiple mating. We present new theory and link it with data to bridge this gap. Two different search protocols exist between species in the bush-cricket genus Poecilimon (Orthoptera): females search for calling males, or males search for calling females. Poecilimon males also transfer a costly nuptial food gift to their mates during mating. We relate variations in searching protocols to variation in nuptial gift size among 32 Poecilimon taxa. As predicted, taxa where females search produce significantly larger nuptial gifts than those where males search. Our model and results show that search roles can reverse when multiple mating brings about sufficiently strong material benefits to females.     

松花江干流大型底栖动物群落结构与水质生物评价

于2010年春季（4-5月）、夏季（7-8月）和秋季（9-11月）,对松花江干流大型底栖动物群落结构进行调查研究,并利用生物指数对松花江干流水质进行评价.共采集到大型底栖动物16目36科116种,其中水生昆虫种类最多,为74种,属6目21科,占总数63.8%,年平均密度为66.80 ind·m^-2、生物量为24.30 g·m^-2.春、夏、秋季的平均密度以春季最高 (90.52 ind·m^-2),秋季(61.26 ind·m^-2)次之,夏季(48.63 ind·m^-2)最低;平均生物量以秋季最高(35.35 g·m^-2),夏季(23.12 g·m^-2)次之,春季(14.41 g·m^-2)最低.Shannon指数、Pielou均匀度指数、Simpson指数均以春季最高,夏季与秋季相近.各断面微生境共有种不多, 物种相似性不高,最大仅为60%;功能摄食群种类数相近,共有撕食者26种,收集者32种,刮食者28种,捕食者30种.采用BI生物指数和FBI生物指数对松花江干流水质的评价结果基本一致,并与化学监测结果基本吻合.松花江干流哈尔滨断面以上水质一般,哈尔滨以下断面水质在不同时期处于污染或严重污染状态.推测大顶子山航电枢纽的修建已对大型底栖动物的物种组成、群落结构造成了较大影响.     

Water status and associated processes mark critical stages in pollen development and functioning

Background

The male gametophyte developmental programme can be divided into five phases which differ in relation to the environment and pollen hydration state: (1) pollen develops inside the anther immersed in locular fluid, which conveys substances from the mother plant – the microsporogenesis phase; (2) locular fluid disappears by reabsorption and/or evaporation before the anther opens and the maturing pollen grains undergo dehydration – the dehydration phase; (3) the anther opens and pollen may be dispersed immediately, or be held by, for example, pollenkitt (as occurs in almost all entomophilous species) for later dispersion – the presentation phase; (4) pollen is dispersed by different agents, remaining exposed to the environment for different periods – the dispersal phase; and (5) pollen lands on a stigma and, in the case of a compatible stigma and suitable conditions, undergoes rehydration and starts germination – the pollen–stigma interaction phase.

Scope

This review highlights the issue of pollen water status and indicates the various mechanisms used by pollen grains during their five developmental phases to adjust to changes in water content and maintain internal stability.

Conclusions

Pollen water status is co-ordinated through structural, physiological and molecular mechanisms. The structural components participating in regulation of the pollen water level, during both dehydration and rehydration, include the exine (the outer wall of the pollen grain) and the vacuole. Recent data suggest the involvement of water channels in pollen water transport and the existence of several molecular mechanisms for pollen osmoregulation and to protect cellular components (proteins and membranes) under water stress. It is suggested that pollen grains will use these mechanisms, which have a developmental role, to cope with environmental stress conditions.     

Fabrication of Biomimetic Water Strider Legs Covered with Setae

Water striders have remarkable water-repellent legs that enable them to stand effortlessly and move quickly on water.Fluidphysics indicates this feature is due to a surface-tension effect caused by the special hierarchical structure of the legs,which arecovered with a large number of inclined setae with fine nanogrooves inducing water resistance.This inspires us to fabricatespecial water-repellent structure on functional surfaces through the cooperation between the surface treatment and the surfacemicro-and nanostructures,which may bring great advantages in a wide variety of applications.In this paper we present aprocedure for fabricating biomimetic water strider legs covered with setae using Polycarbonate Track-Etched(PCTE)membranesas templates.By choosing appropriate membrane lengths,diameters,pitches and densities of the setae,the biomimeticlegs can be fabricated conveniently and at a low cost.Furthermore we investigated the relationship between stiffness of themolding materials,high aspect ratio and density,which affect the fidelity of fabrication and self adhesion,to optimize thestability of setae.The knowledge we gained from this study will offer important insights into the biomimetic design and fabricationof water strider setae.     

Root pressure and specific conductivity in temperate lianas: exotic Celastrus orbiculatus (Celastraceae) vs. native Vitis riparia (Vitaceae)

The exotic temperate liana (woody vine) Celastrus orbiculatus has become a weed in Michigan, occurring in many of the same habitats as the native liana Vitis riparia. However, C. orbiculatus frequently develops into extensive monospecific infestations, while V. riparia does not. Freezing-induced embolism may be responsible for limiting liana distribution. Root pressure has been observed in numerous tropical lianas and temperate species of Vitis and has been implicated as vital to the recovery of xylem function in wide vessels following winter freezes. For both of these co-occurring lianas we investigated root pressure and water conductance as possible explanatory factors for their differential spread. According to our hypothesis, C. orbiculatus should have produced greater or more frequent root pressures than V. riparia. However, the reverse proved true, indicating that root pressure is not a prerequisite for weedy proliferation of C. orbiculatus. Additionally, the seasonal patterns of specific conductivity of stem xylem indicate that each species responds differently to environmental constraints. Vitis riparia establishes conductivity early in the growing season, before the leaves emerge, using root pressure to reverse embolism, but loses conductivity with the first freeze in early autumn. Celastrus orbiculatus is slow to establish conductivity, depending on new wood production, but leafs out sooner than V. riparia and maintains green leaves after the first freeze. Vulnerability curves of xylem to cavitation caused by water stress for the two species indicate that they respond similarly to dehydration. These results indicate that root pressures are not responsible for the invasive success of C. orbiculatus and suggest that other factors must be key to its prolific invasion.     

植物根系吸水机理的研究进展

近年来,植物根系吸水机理在细胞、组织和整体水平上的研究进展非常迅速,对阐明植物抗旱机制及其高效利用有限水资源途径的探讨具有重要意义.本文主要对植物根的复合结构和根系在土壤中的分布、根系中水流性质等方面的最新研究状况进行了概述,特别详细地论述了水通道蛋白的表达及功能与根系中水分运动的关系、以及根系输水的调节和根系吸水过程中的信号传导方面的研究动态,并且评价了根的复合运输模型和根系吸水的数学模型等,最后就其可能生理意义及其应用前景作了评述.