Phagotrophic Protist Diversity in the Groundwater of a Karstified Aquifer – Morphological and Molecular Analysis

To clarify the structure of microbial food webs in groundwater, knowledge about the protist diversity and feeding strategies is essential. We applied cultivation‐dependent approaches and molecular methods for further understanding of protist diversity in groundwater. Groundwater was sampled from a karstified aquifer located in the Thuringian Basin (Thuringia, Germany). Cultivable protist abundance estimated up to 8,000 cells/L. Eleven flagellates, 10 naked amoebae, and one ciliate morpho‐species were detected in groundwater enrichment cultures. Most of the flagellates morpho‐species, typically < 10 μm, were sessile or free swimming suspension feeders, e.g., Spumella spp., Monosiga spp., and mobile, surface‐associated forms that grasp biofilms, e.g., Bodo spp. Naked amoebae, typically < 35 μm, that grasp biofilms were represented by, e.g., Vahlkampfia spp., Vannella spp., and Hartmanella spp. The largest fraction of the 18S rRNA gene sequences was affiliated with Spumella‐like Stramenopiles. Besides, also sequences affiliated with fungi and metazoan grazers were detected in clone libraries of the groundwater. We hypothesize that small sized protist species take refuge in the structured surface of the fractures and fissures of the karstified aquifer and mainly feed on biofilm‐associated or suspended bacteria.     

Biodeterioration of asbestos cement (AC) pipe in drinking water distribution systems

Various types of microorganisms have been found to inhabit the inner surfaces of asbestos cement (AC) pipe and their activities can cause significant structural damage. They cause a patina to form on the inside surface of AC pipes as a distinctively continuous coating, commonly 2-5 mm in thickness and generally pigmented as yellow, orange, brown or black depending on the metallic cations that have been incorporated into the surface of biofilm (bioaccumulation). Four sublayers can be identified in the patina, from the outer sublayer that directly interacts with the conveyed drinking water to the inner sublayer that is in proximity of the intact cement matrix. The microbes in the outer sublayer are composed mainly of inactive biomass that separates the aerobic environment of the flowing water from the anaerobic conditions inside the patina. The bacteriological community structure shifts from mixed heterotrophic bacteria (HAB), iron-related bacteria (IRB) and slime-forming bacteria (SLYM) in the outer layer, to a more diverse community with IRB, acid-producing bacteria (APB) and SLYM and HAB in the middle sublayer, and further to the SLYM dominated in the inner sublayer. By directly interacting with cementitious materials, including generating organic acids, IRB and APB play important roles in the leaching of free lime and the dissolution of calcium (Ca)-bearing hydrated components of AC pipes, creating porous structure and reducing the pipe strength. Scanning electron microscopy with an energy dispersive X-ray has revealed that bacterial activity on the internal AC pipe wall had resulted in a significant loss of hydrated cement matrix, which can cause pipe failure when stresses imposed on the pipe exceed the remaining pipe strength.     

The unicellular freshwater cyanobacterium Synechococcus and mixotrophic flagellates: evidence for a functional association in an oligotrophic, subalpine lake

1. We applied Reynolds's approach to the study of phytoplankton ecology through functional associations of species to identify possible algal species, which associate with the freshwater cyanobacterium Synechococcus. Previously an association among Synechococcus spp. and small‐celled chlorophytes (association Z) has been recognised by evaluating phytoplankton associations according to functional criteria. 2. Biomass data for phytoplankton and picocyanobacteria from Lago Maggiore spanning more years were organised in a matrix and a cluster analysis was performed. The results showed four groups separated at a linkage distance of 0.20. Mixotrophic species which clustered with Synechococcus spp. were Ceratium hirundinella, Chrysochromulina parva, Cryptomonas erosa, Cryptomonas ovata, Dinobryon bavaricum, Dinobryon sociale, Rhodomonas minuta and Uroglena americana. The redundancy analysis (RDA) consolidated the association of Synechococcus with C. hirundinella and R. minuta showing greater probability of occurrence than random aggregates of species. 3. The association Synechococcus spp. –C. hirundinella also appeared from temporal variation of their biomass. In early summer both these species increased at the same time; later, the peak of C. hirundinella (potential predator) coincided with a minimum of Synechococcus spp. suggesting a possible predator‐prey interaction. This implied that phytoplankton assemblages which form a functional group cannot only have similar adaptations and requirements but can also exhibit trophic interactions. 4. We propose to enlarge the association Z and create an association Z_MX (where _MX stands for mixotrophs) which would include Synechococcus spp. and C. hirundinella as the most representative of the mixotrophic species found in the oligotrophic Lago Maggiore.     

ORIENTATION OF SWIMMING FLAGELLATES BY SIMULTANEOUSLY ACTING EXTERNAL FACTORS1

The directionality of phototaxis combined with gravitaxis was investigated experimentally for populations of the swimming alga Euglena gracilis Klebs. Two irradiances were used: a “weak” irradiance to elicit positive phototaxis and a “strong” irradiance to elicit negative phototaxis. In addition, by changing the density of cells in the suspension, the number of collisions between cells was varied to determine the effects of these collisions on the distribution of swimming directions in both the absence and the presence of illumination. We found that positive phototaxis was associated with a broader distribution of swimming directions than was negative phototaxis. In the latter case, the effect of phototaxis dominated over that of gravitaxis. Experiments on another swimming alga, Chlamydomonas nivalis Wille, showed that collisions between cells degraded the directionality of gravitaxis.     

Temperature-shock induction of multiple flagella induces additional synthesis of flagellum specific mRNAs and tubulin

Four mRNAs (alpha- and beta-tubulin, flagellar calmodulin and Class-I), specifically expressed when Naegleria amebae differentiate into flagellates, were followed at 5-10 min intervals during the temperature-shock induction of multiple flagella in order to better understand how basal body and flagellum number are regulated. Surprisingly, tubulin synthesis continued during the 37 min temperature shock. An initial rapid decline in alpha- and beta-tubulin and flagellar calmodulin mRNAs was followed by a rapid re-accumulation of mRNAs before the temperature was lowered. mRNA levels continued to increase until they exceeded control levels by 4-21%. Temperature shock delayed flagella formation 37 min, produced twice as much tubulin protein synthesis and three fold more flagella. Labeling with an antibody against Naegleria centrin suggested that basal body formation was also delayed 30-40 min. An extended temperature shock demonstrated that lowering the temperature was not required for return of mRNAs to near control levels suggesting that induction of multiple flagella and the formation of flagella per se are affected in different ways. We suggest that temperature-shock induction of multiple flagella reflects increased mRNA accumulation combined with interference with the regulation of the recently reported microtubule-nucleating complex needed for basal body formation.     

Effects of digenean trematodes and heterotrophic bacteria on mortality and burying capability of the common cockle Cerastoderma edule (L.)

Population dynamics of marine invertebrates is controlled by a variety of abiotic and biotic factors. Among these, some have received lesser attention from marine ecologists because of their ‘discrete’ nature. This is the case of parasitism and bacterial load. In the present study, we focused on the role that both digenean trematodes and heterotrophic aerobic bacteria might play in the mortality and burying behaviour of cockles, Cerastoderma edule.The bivalves were sampled monthly during 1 year from two sites in Arcachon Bay (French Atlantic coast). Mortality rates were assessed after transferring in the laboratory normally buried and unburied (i.e. found lying at the sediment surface at low tide) cockles. Their digenean and bacterial loads were determined for both positions (normally buried and unburied). Mortality rate was significantly higher for cockles found out of the sediment at low tide, suggesting that this abnormal position was a prelude to cockles' death. Comparison of digenean load of cockles showed no significant difference between buried and unburied bivalves. In contrast, bacterial load was significantly higher in unburied cockles than in normally buried animals. The effect of high concentration of a marine bacterial strain (Pseudomonas fluorescens) on cockles' burying behaviour and mortality was tested in the laboratory. Results showed that these bacteria could trigger the emergence of animals from the sediment but did not cause cockles' death. These field observations and laboratory experiments suggest that bacteria, rather than digenean trematodes, could play a role in the emergence of cockles and, hence, affect their survival in the wild.     

泥螺养殖滩涂异养菌群和弧菌的检测

1　引　　言泥螺 (Ｂｕｌｌａｃｔａｅｘａｒａｔａ)隶属腹足纲软体动物 ,肉质鲜嫩、营养丰富 ,是浙江沿海重要的滩涂养殖品种 .至 2 0 0 0年浙江省养殖面积已达 1.13× 10 4ｈｍ2 ,由于实行生态养殖 ,本轻利厚 ,广受养农的青睐 .随着泥螺养殖生产的发展 ,养殖业的自身污染和陆源排污水的大量入海 ,养殖生态环境越趋恶化 ,使生物病原大量滋生 ,自 1995年以来常诱发养殖泥螺的暴发性死亡 .有关海洋滩涂环境细菌的研究至今未见报道 .为探明泥螺养殖周期内暴发性死亡与滩涂环境的关系 ,对养殖滩涂的细菌菌群组成及其生态特性进行了检测分析 .2…     

Cospeciation of termite gut flagellates and their bacterial endosymbionts: Trichonympha species and 'Candidatus Endomicrobium trichonymphae'

Symbiotic flagellates play a major role in the digestion of lignocellulose in the hindgut of lower termites. Many termite gut flagellates harbour a distinct lineage of bacterial endosymbionts, so-called Endomicrobia, which belong to the candidate phylum Termite Group 1. Using an rRNA-based approach, we investigated the phylogeny of Trichonympha , the predominant flagellates in a wide range of termite species, and of their Endomicrobia symbionts. We found that Trichonympha species constitute three well-supported clusters in the Parabasalia tree. Endomicrobia were detected only in the apical lineage (Cluster I), which comprises flagellates present in the termite families Termopsidae and Rhinotermitidae, but apparently absent in the basal lineages (Clusters II and III) consisting of flagellates from other termite families and from the wood-feeding cockroach, Cryptocercus punctulatus . The endosymbionts of Cluster I form a monophyletic group distinct from many other lineages of Endomicrobia and seem to have cospeciated with their flagellate host. The distribution pattern of the symbiotic pairs among different termite species indicates that cospeciation of flagellates and endosymbionts is not simply the result of a spatial separation of the flagellate lineages in different termite species, but that Endomicrobia are inherited among Trichonympha species by vertical transmission. We suggest extending the previously proposed candidatus name ' Endomicrobium trichonymphae ' to all Endomicrobia symbionts of Trichonympha species, and estimate that the acquisition by an ancestor of Trichonympha Cluster I must have occurred about 40–70 million years ago, long after the flagellates entered the termites.