Hydrolysis of aromatic beta-glucosides by non-pathogenic bacteria confers a chemical weapon against predators

Bacteria present in natural environments such as soil have evolved multiple strategies to escape predation. We report that natural isolates of Enterobacteriaceae that actively hydrolyze plant-derived aromatic beta-glucosides such as salicin, arbutin and esculin, are able to avoid predation by the bacteriovorous amoeba Dictyostelium discoideum and nematodes of multiple genera belonging to the family Rhabditidae. This advantage can be observed under laboratory culture conditions as well as in the soil environment. The aglycone moiety released by the hydrolysis of beta-glucosides is toxic to predators and acts via the dopaminergic receptor Dop-1 in the case of Caenorhabditis elegans. While soil isolates of nematodes belonging to the family Rhabditidae are repelled by the aglycone, laboratory strains and natural isolates of Caenorhabditis sp. are attracted to the compound, mediated by receptors that are independent of Dop-1, leading to their death. The b-glucosides-positive (Bgl(+)) bacteria that are otherwise non-pathogenic can obtain additional nutrients from the dead predators, thereby switching their role from prey to predator. This study also offers an evolutionary explanation for the retention by bacteria of `cryptic' or `silent' genetic systems such as the bgl operon.

Food and feeding habits of Heterotis niloticus from River Kaduna floodplain

A total of 61 Heterotis niloticus specimen were examined and evaluated to determine the food and feeding habits using the frequency of occurrence, point and dominance methods. The specimens had mean standard length of 27.09 plus or minus 4.73cm, total length of 33.40cm body weight of 90-900g. The gut length ranges from 34-104cm while the gut weight range from 2.79-130g. It was observed that the fish fed mostly on plankton with rotifers and polyscysits having the highest number of frequency and dominant value with mean value of 43.03 plus or minus 4.12 and 11.73 plus or minus 1.15, 37.45 plus or minus 3.27 and 8.32 plus or minus 0.38 respectively. Arcella had the least mean frequency of occurrence of 3.27 plus or minus 17. Amoeba sp had the least mean dominance value of 7.06 plus or minus 50 and Aphnocapsa sp had the least mean frequency of occurrence of 1.10 plus or minus 0.29 and Navicula sp had the least mean dominance value of 4.31 plus or minus 1.11. Heterotis niloticus of River Kaduna flood plain is therefore considered to be predominantly planktivorous

Part I. Regulation of development in the cellular slime mold Dictyostelium discoideum. Part II. Polysomes and RNA synthesis during early development of the surf clam Spisula solidissima

Part I. The cellular slime mold Dictyostelium discoideum is a simple eukaryote which undergoes a multi-cellular developmental process. Single cell myxamoebae divide vegetatively in the presence of a food source. When the food is depleted or removed, the cells aggregate, forming a migrating pseudoplasmodium which differentiates into a fruiting body containing stalk and spore cells. I have shown that during the developmental cycle glycogen phosphorylase, aminopeptidase, and alanine transaminase are developmentally regulated, that is their specific activities increased at a specific time in the developmental cycle. Phosphorylase activity is undetectable in developing cells until mid-aggregation whereupon it increases and reaches a maximum at mid-culmination. Thereafter the enzyme disappears. Actinomycin D and cycloheximide studies as well as studies with morphologically aberrant and temporally deranged mutants indicate that prior RNA and concomitant protein synthesis are necessary for the rise and decrease in activity and support the view that the appearance of the enzyme is regulated at the transcriptional level. Aminopeptidase and alanine transaminase increase 3 fold starting at starvation and reach maximum activity at 18 and 5 hours respectively.

The cellular DNA s of D. discoideum were characterized by CsC1 buoyant density gradient centrifugation and by renaturation kinetics. Whole cell DNA exhibits three bands in CsCl: ρ = 1.676 g/cc (nuclear main band), 1.687 (nuclear satellite), and 1.682 (mitochondrial). Reassociation kinetics at a criterion of Tm -23°C indicates that the nuclear reiterated sequences make up 30% of the genome (Cot_1/2 (pure) 0.28) and the single-copy DNA 70% (Cot_1/2(pure) 70). The complexity of the nuclear genome is 30 x 10⁹ daltons and that of the mitochondrial DNA is 35-40 x 10⁶ daltons (Cot_1/2 0.15). rRNA cistrons constitute 2.2% of nuclear DNA and have a ρ = 1.682.

RNA extracted from 4 stages during developmental cycle of Dictyostelium was hybridized with purified single-copy nuclear DNA. The hybrids had properties indicative of single-copy DNA-RNA hybrids. These studies indicate that there are, during development, qualitative and quantitative changes in the portion of the single-copy of the genome transcribed. Overall, 56% of the genome is represented by transcripts between the amoeba and mid-culmination stages. Some 19% are sequences which are represented at all stages while 37% of the genome consists of stage specific sequences.

Part II. RNA and protein synthesis and polysome formation were studied during early development of the surf clam Spisula solidissima embryos. The oocyte has a small number of polysomes and a low but measurable rate of protein synthesis (leucine-³H incorporation). After fertilization, there is a continual increase in the percentage of ribosomes sedimenting in the polysome region. Newly synthesized RNA (uridine-5-³H incorporation) was found in polysomes as early as the 2-cell stage. During cleavage, the newly formed RNA is associated mainly with the light polysomes.

RNA extracted from polysomes labeled at the 4-cell stage is polydisperse, nonribosomal, and non-4 S. Actinomycin D causes a reduction of about 30% of the polysomes formed between fertilization and the 16-cell stage.

In the early cleavage stages the light polysomes are mostly affected by actinomycin.

Morphology, biometry and distribution of Difflugia biwae Kawamura, 1918 (Protozoa : Rhizopoda)

The freshwater testate amoeba Difflugia biwae Kawamura, 1918, isolated from Mulan Lake, Hubei Province, China is investigated using light microscopy and scanning electron microscopy. The morphology, biometry and distribution of this little known species are supplied. After careful comparison with three other similar species, including D. delicatula Gauthier-Livre et Thomas, D. elegans Penard and D. oblonga caudata Stepanek, we believe that the characteristics of smooth fusiform shell, conspicuous great collar flare (always larger than body-width) around the aperture, constriction behind the aperture and a somewhat curved aboral horn set D. biwae apart from all other Difflugia species. This species shows a great diversity in total length, collar height and aboral horn length which have high variability (CV between 11.76 and 24.52). However, body width, collar diameter, neck width, body length and aperture diameter are fairly constant with low variability (CV between 5.34 and 8.79) which shows a remarkable uniformity of D. biwae. Also, the size frequency distributions of both body width and body length yield bell-shaped (normally distributed) curves and indicate that D. biwae is a size-monomorphic species, characterized by a main-size class and a small size range. D. biwae is probably endemic to East Asia (China and Japan) because it has such a large size (165-306 mu m) that it would have been easily found in Europe and North America, if it were there. Consequently, D. biwae must have a restricted geographical distribution, disproving the old hypothesis that microscopic organisms are cosmopolitan.

Redescription of Difflugia tuberspinifera Hu, Shen, Gu et Gong, 1997 (Protozoa : Rhizopoda : Arcellinida : Difflugiidae) from China

The freshwater testate amoeba Difflugia tuberspinifera Hu et al. 1997 collected from pond and lake in China, is investigated by light and scanning electron microscopy. This little known taxon is redescribed and its morphology, biometry and ecology are supplied. After carefully comparison with other six similar species including Difflugia bartosi Stepanek, D. corona Wallich, D. corona cashi Deflandre, D. corona tuberculata Vucetich, D. muriformis Gauthier-Lievre et Thomas and Netzelia tuberculata (Wallich) Netzal we believe that the sub-spherical to spherical shell, the mulberry-shaped appearance, the 7-10 apertural tooth-like structures, the short collar and the conical spines numbering from 4 to 8 at the upper equatorial region in D. tuberspinifera set it apart from other species. Besides, statistical analysis indicates that D. tuberspinifera is a size-monomorphic species characterized by a main-size class and a small size range and the shell height is significant correlated with other morphometric characters at p < 0.05 excepting the number of aperture tooth-like structures and the number of spines. Moreover, D. tuberspinifera inhabits not only lotic but also lentic environment.

一种可变形移动机器人协同构形变换方法

提出了一种可变形移动机器人AMOEBA-I的协同构形变换方法,建立了机器人系统的数学模型,对各个模块之间的协同变换及运动特性进行了分析.研究了机器人3个模块在协同变换过程中的电流变化情况,实现了3种特殊构形之间的变换.通过理论分析和实验比较了协同构形变换方法的特点,实验验证了在多种地面条件下机器人协同构形变换方法的有效性.

基于网络的自重构机器人重构路径研究

文中针对自重构机器人的重构问题提出了一种基于网络的分析方法.自重构模块机器人"AMOEBA-I"的9种不同构形组成了一个构形网络.机器人的每种构形被看成带有权值的有向构形网络中的一个节点.一种构形向另一种构形的转换可描述为一条非负值的有向路径.将图论应用于构形变化的分析,根据构形的拓扑信息相应定义了重构路径、可重构矩阵和路径矩阵.在此基础上,将图论中的算法应用于重构路径的计数和最佳重构路径的选择.数值分析与仿真实验结果验证了该方法的可行性.同时,该方法还可以用于其他自重构机器人的构形控制与自重构规划.

可变形机器人协同转向运动研究

可变形机器人AMOEBA-I具有多种构形和多种转向方式.为实现机器人转向性能的优化,提出了可变形机器人的协同转向方法,并建立了相应的数学模型,对不同构形下的协同转向方式进行了理论分析.设定了机器人三个模块在协同转向过程中的运动关系,在此基础上给出了可变形机器人协同转向性能的评价指标.通过理论和实验比较了不同构形下的协同转向方式,实验验证了协同转向方法的有效性.

可变形机器人协同越障方法及控制策略研究

在复杂环境中工作的机器人对越障能力的要求越来越高。为了充分发挥可变形机器人AMOEBA-Ⅰ的越障能力,增强机器人在非结构环境中的适应性,提出了可变形机器人的协同越障方法。建立了数学模型,对机器人越障高度与其重心位置的关系进行了理论分析。从理论上比较了常规越障方法与协同越障方法所能翻越障碍的最大高度。同时提出了自主越障过程的控制策略,采用了根据情感状态的变化对机器人控制策略进行微调,并建立了相应地情感模型。实验验证了协同越障方法及自主越障控制策略的有效性。

可变形机器人自适应路径规划研究

可变形机器人AMOEBA-Ⅰ具有多种构形方式,通过构形的改变可以增强机器人在狭窄空间中的通过能力.本文提出了将可变形机器人的变形能力融入到势场法中进行自适应路径规划的方法,以充分发挥可变形机器人特有的通过性能.研究了改进势场法,有效地解决了局部极小问题。引入了可见度概念,研究了减小计算量提高实时性品质指标的方法。通过实验结果证明机器人能够根据环境变化改变自身构形,表现自适应路径规划.同时有效地缩短了路径。

一种可重构模块机器人中心构形的选择方法

可重构模块机器人具有多种构形以适应不同环境和任务的要求,构形的多变增加了构形研究的难度.在可重构模块机器人的众多构形中,中心构形作为可重构模块机器人的首选构形或基准构形,对系统的实际应用有重要参考价值.文中提出了一种在所有构形中选择一个中心构形的方法.根据构形之间可以相互转化的拓扑特征,利用网络图中的基本思想和原理对可重构模块机器人的构形进行建模;相应定义了构形转换耗值矩阵和构形中心因子,根据最大构形中心因子可以对中心构形进行选择.以中国科学院沈阳自动化研究所研制的三模块可重构机器人AMOEBA-I为例,利用仿真计算的结果对机器人9种构形的中心因子进行计算和比较,验证了该方法的可行性.最后根据构形邻接数,给出了中心构形选择方法的应用举例.此方法还可以适用于其他可重构模块机器人系统中心构形的选择.

Relacje między zgrupowaniami ameb skorupkowych, roślinnością i parametrami siedliskowymi torfowisk minerotroficznych Wielkopolski

Wydział Biologii: Instytut Biologii Środowiska/Zakład Hydrobiologii

Molecular analysis of virulence mechanisms associated with adherent-invasive Escherichia coli (AIEC)

Crohn's Disease (CD) is a chronic inflammatory bowel disease of unknown etiology. Recent work has shown that a new pathotype of Escherichia coli, Adherent Invasive E. coli (AIEC) may be associated with CD. AIEC has been shown to adhere to and invade epithelial cells and to replicate within macrophages (together this is called the AIEC phenotype). In this thesis, the AIEC phenotype of 84 E. coli strains were determined in order to identify the prevalence of this phenotype within the E. coli genus. This study showed that a significant proportion of E. coli strains (approx. 5%) are capable of adhering to and invading epithelial cells and undergoing intramacrophage replication. Moreover, the results presented in this study indicate a correlation between survival in macrophage and resistance to grazing by amoeba supporting the coincidental evolution hypothesis that resistance to amoebae could be a driving force in the evolution of pathogenicity in some bacteria, such as AIEC. In addition, this study has identified an important regulatory role for the CpxA/R two component system (TCS) in the invasive abilities of AIEC HM605, a colonic mucosa-associated CD isolate. A mutation in cpxR was shown to be defective in the invasion of epithelial cells and this defect was shown to be independent of motility or the expression of Type 1 fimbriae, factors that have been shown to be involved in the invasion of another strain of AIEC, isolated from a patient with ileal CD, called LF82. The CpxA/R TCS responds to disturbances in the cell envelope and has been implicated in the virulence of a number of Gram negative pathogens. In this study it is shown that the CpxA/R TCS regulates the expression of a potentially novel invasin called SinH. SinH is found in a number of invasive strains of E. coli and Salmonella. Moreover work presented here shows that a critical mechanism underpinning AIEC persistence in macrophages is the repair of DNA bases damaged by macrophage oxidants. Together these findings provide evidence to suggest that AIEC are a diverse group of E. coli and possess diverse molecular mechanisms and virulence factors that contribute to the AIEC phenotype. In addition, AIEC may have gone through different evolutionary histories acquiring various molecular mechanisms ultimately culminating in the AIEC phenotype. The gastrointestinal (GI) tract harbors a diverse microbiota; most are symbiotic or commensal however some bacteria have the potential to cause disease (pathobiont). The work presented here provides evidence to support the model that AIEC are pathobionts. AIEC strains can be carried as commensals in healthy guts however, when the intestinal homeostasis is disrupted, such as in the compromised gut of CD patients, AIEC may behave as opportunistic pathogens and cause and/or contribute to disease by driving intestinal inflammation.

Testate amoebae as indicators of hydroseral change: an 8,500 year record from Mer Bleue Bog, eastern Ontario, Canada.

Testate amoebae have been used widely as a proxy of hydrological change in ombrotrophic peatlands, although their response to abiotic controls in other types of mire and fenland palaeo-environments is less well understood. This paper examines the response of testate amoebae to hydroseral and other environmental changes at Mer Bleue Bog, Ontario, Canada, a large ombrotrophic peatland, which evolved from a brackish-water embayment in the early Holocene. Sediments, plant macrofossils and diatoms examined from a 5.99 m core collected from the dome of the bog record six stages of development: i) a quiet, brackish-water riverine phase (prior to ca. 8500 cal BP); ii) a shallow lake (ca. 8500–8200 cal BP); iii) fen (8200–7600 cal BP); iv) transitional mire (7600–6900 cal BP); v) pioneer raised mire (6900–4450 cal BP); and vi) ombrotrophic bog (4450 cal BP-present).

Testate amoebae, notably small (<25 µm diameter) specimens of Centropyxis aculeata type, first appear in low abundances in sediments ascribed to the lacustrine phase. Diatoms from the same horizons record a shallowing in water depth, a decline in salinity and the development of emergent macrophytic vegetation, which may have provided favourable conditions for testate amoeba colonization. The testate amoeba communities of the inferred fen phase are more diverse and include centropyxids, cyclopyxids, Arcellidae and Hyalospheniidae, although the assemblages show some differences to those recently reported in modern European fen environments. The Fen–Bog Transition (FBT) is also dominated by C. aculeata type. The change in testate amoeba communities around this key transition is apparent in the results of Detrended Correspondence Analysis (DCA), and appears to reflect a latent nutrient gradient and a secondary moisture gradient. DCA analyses of plant macrofossil remains around the FBT show a similar trend, although the sensitivity of the two proxies to the inferred environmental changes differs. Comparisons with other regional mid-Holocene peatland records confirm the important influence of reduced effective precipitation on the testate amoeba communities during the initiation and development of Sphagnum-dominated, raised bog communities.

Mid to late Holocene hydrological change in continental eastern Canada: assessing regional trends from ombrotrophic peat records.

ABSTRACT High resolution records of mid-late Holocene hydro-climatic change are presented from Mer Bleue Bog, eastern Ontario. Past climatic changes in this region have previously been inferred from lake sediments, but rain-fed peatlands can offer additional insights into the spatial and temporal pattern of moisture availability. In this study, reconstructed water table depths are based on a testate amoeba-derived transfer function developed for the region and changes in bog surface wetness are compared with plant macrofossil and peat humification data.

RÉSUMÉ Nous présentons les enregistrements hautes résolutions des variations hydrologique durant la second moitié de l’Holocène pour les tourbières Mer Bleue á l’est de l'Ontario. Précédemment, les changements climatiques de cette région ont été dérivés à partir de prélèvement de sédiments de lac. Mais ils s’avèrent que les tourbières ombrotrophes offrir un éclairage supplémentaire sur les schémas de répartition spatiale et temporelle de la disponibilité de l'humidité. Dans cette étude, des profondeurs reconstruites de nappe phréatique sont basées sur un modèle de function de transfert d’amibes (Arcellinida) et des changements de l’humidité de surface de la tourbière sont comparés avec les macrofossils et au humification de tourbe dans une analyse multi-proxy.