Influence possible des protozoaires sur le taux de mortalité des bactéries autotrophes nitrifiantes

Le modèle de l'IAWQ du processus de boues activées représente les mécanismes endogènes de la biomasse nitrifiante par le décès des micro-organismes (équation d'ordre 1 par rapport à la biomasse). La constante de décès, ou taux de mortalité bA, est aujourd'hui encore mal connue, et en particulier les facteurs influants sur sa valeur. De récentes études ont montré que la prédation par la microfaune pourrait être un facteur déterminant sur la valeur de bA. Cette étude se propose donc de quantifier l'effet de la prédation sur la valeur de bA. Deux réacteurs maintenus sans alimentation en substrat ont été caractérisés en parallèle: l'un a reçu une dose d'antibiotique spécifique aux eucaryotes (cycloheximide) afin de diminuer la quantité d'organismes de la microfaune, alors que l'autre n'a reçu aucun antibiotique (témoin). Les résultats obtenus montrent que le cycloheximide inhibe la plupart des organismes de la microfaune sauf les amibes; celles-ci semblent plutôt stimulées par cet antibiotique. En ce qui concerne la nitrification, un ralentissement de la production de nitrate dans le réacteur traité à l'antibiotique est observé à partir du sixième jour. Cette diminution de production de nitrate est probablement causée par une réduction de l'azote nitrifiable (qui est mobilisé par les amibes) couplée à une prédation des organismes nitrifiants par les amibes. D'ailleurs, l'augmentation de la prédation par les amibes à partir du jour 6 a diminué l'activité nitrifiante également mesurée par respirométrie (rO2 Nmax). Cette diminution du taux de respiration indique une augmentation du taux de mortalité (bA) des organismes nitrifiants. En effet, la valeur du taux de mortalité mesurée dans le réacteur témoin est de 0.08 d-1 alors que selon la microfaune présente dans le réacteur inhibé au cycloheximide, la valeur de ce taux de mortalité a varié entre 0.05 d-1 et 0.15 d-1.Designing biological wastewater treatment plants with the aid of the model developed by the IAWQ requires the knowledge of biological kinetic parameters. For nitrifying activated sludge, these parameters are related to nitrifying bacteria: maximum autotrophic growth rate µAmax, yield coefficient YA and the autotrophic decay rate bA. Although variables influencing µAmax and YA values are well known, this is not the case for bA. MARTINAGE and PAUL (2000) have recently shown that the bA value is strongly influenced by the influent quality, leading to the assumption that influent quality has a strong effect on microfauna composition, and consequently on the grazing rate of microfauna on nitrifying bacteria. In fixed-film processes, protozoan grazing reduces the bacterial population considerably (NATUSCKA and WELANDER, 1994). However, although many data are available concerning the grazing rates of different protozoa, the effect of microfaunal grazing on nitrification is still a matter of debate (RATSAK et al., 1994) and its effect on the bA value is still unknown. These two topics are investigated here.Nitrifying activated sludges were grown in two identical batch reactors, but in one, cycloheximide was added to inhibit eucaryotic growth (MAURINES CARBONEILL et al., 1998). Microfauna organism numbers were quantified in both reactors by microscopic observations of flagellated protozoa (>8 µm), amoebae, ciliates, rotifers and higher invertebrates (Fig. 3). Microbial counts were then correlated with the bA value. The latter was determined using the procedure proposed by SALZER (1992) which consists of characterising the time behaviour of the maximum nitrification rate measured by respirometry of activated sludge under substrate starvation. Under these conditions bacteria die and organic nitrogen is released into the bulk phase. This nitrogen is ammonified, and nitrifying bacteria use this substrate to produce nitrate, and then autotrophic bacterial growth occurs. This method takes this growth into account by characterising nitrate production during the experiment (Fig. 2).The effect of cycloheximide on nitrification was first determined to make sure that this compound is not inhibitory toward nitrifiers. Results obtained (Table 1) show that cycloheximide was not inhibitory toward nitrate production or the maximum nitrification oxygen uptake rate (rO2 N) after 4 hours of contact with nitrifying biomass. Cycloheximide addition in the activated sludge had an important impact on rotifers and flagellates but no effect on ciliates; it also seemed to stimulate amoebae growth. In both reactors, flagellates were mainly Peranama, attached ciliates were mainly Opercularia and Epistylis and a few Vorticella. Free ciliates like Aspidisca and Euplotes were found in both reactors.Variation with time of the abundance of microfauna organisms is shown in Figures 4 and 5 for both reactors. In the reference reactor the number of microfauna organisms decreased with time (Fig. 4) probably due to substrate starvation. Microfauna composition remained however diversified. For the inhibited reactor (Fig. 5), three periods were observed. During period I, the microfauna was mainly composed of ciliates and the number of microfaunal organisms decreased rapidly. During period II, an important growth of amoebae was observed. Cycloheximide was then added during this period to reduce their number. This growth of amoebae seems to be caused by the resistance of these micro-organisms toward inhibiting compounds (SRIKANTH et BERK, 1993). During period III, the number of microfaunal organisms was lower than during period II, and microfauna was mainly composed of ciliates.Nitrate concentration behaviour, necessary for bA calculation, is shown on Figure 6. In the reference reactor, nitrate concentrations varied linearly. For the inhibited reactor, the linear pattern was not observed during period II. This result was probably caused by an important nitrogen assimilation need of amoebae (ELDRIGE and JACKSON, 1993). Because organic nitrogen released by bacterial decay is consumed by amoebae assimilation, less nitrogen is available for the ammonification process and therefore for nitrification. Ammonia concentrations remained below 0.2 mg N·l-1 during all the experiment for both reactors. When amoebae disappeared from the inhibited reactor (period III) nitrate concentration varied linearly again.Variations of the maximum nitrification oxygen uptake rate (rO2 Nmax) with time are presented in Figure 7 (A&B) for both reactors. Two curves are plotted on each figure. Empty squares represent the measured rO2 N and black points represent the maximum nitrification rate that would have been measured if no growth on ammonification products had occurred. For the reference reactor (Fig. 7A), a value for bA of 0.08 d-1 can be calculated and can be considered constant for a constant microfauna composition.Three bA values can be estimated for the reactor inhibited with cycloheximide (Fig. 7B), corresponding to the three periods observed for microfauna composition. During period I, the bA value is 0.05 d-1 : a decrease in the microfaunal organism numbers implies a decrease of the bA value. During period II, when a development of amoebae is observed, the bA value increases and reaches 0.15 d-1. During period III with reduced grazing, the bA value is 0.13 d-1. Since during periods I and III the microfauna is mainly composed of ciliates, this difference between bA values is likely due to the observed difference in floc size between periods I and III.The results obtained during this study tend to prove (1) that the use of cycloheximide reduces microfaunal populations but can lead to a development of amoebae, and (2) that microfauna grazing seems to have an influence on the bA value, which can vary from 0.05 to 0.15 d-1 depending on microfaunal composition and abundance

Novel Pausing Behavior Of The Kinesin-3 Family Member KIF1A Is Regulated By Tau

The unique relationship between neuronal structure and function is paramount for the complexity of the human nervous system. This relationship allows neurons to receive, process, and transmit information through many inter- and intracellular mechanisms. Axonal transport is an essential intracellular mechanism for neuronal health and viability. This process involves the transport of cellular cargo in the anterograde and retrograde direction along the axon, relaying materials between the soma and axon terminals, respectively. The necessity of an expedited form of transport becomes clear when one considers the magnitude of distance cargo must travel; in some cases, the axon can be up to a meter in length. While axonal transport increases the efficiency of cargo delivery, it is imperative that this process be regulated both spatially and temporally. The choreography of axonal transport is mediated by molecular motor proteins that carry neuronal cargo along microtubule tracks within the axon. Kinesin proteins, a class of molecular motors, transport cargo in the anterograde direction. KIF1A is a kinesin-3 family member that is responsible for the transport of certain critical intracellular cargo. However, the mechanisms for KIF1A regulation remain largely misunderstood. A known regulatory mechanism of kinesin motors is via the presence of microtubule associated proteins (MAPs) that bind to and crowd microtubule roadways. Specifically, the neuronal MAP Tau has been shown to differentially regulate kinesin families found in the neuron, such as kinesin-1 and kinesin-2. Furthermore, these regulatory capabilities are directly related to the behavioral binding state of Tau, of which it can bind statically or diffusively to the microtubule. While the potential regulatory relationship between KIF1A and Tau is unknown, both of these players exhibit pathological behavior in neurodegenerative diseases such as Alzheimer’s disease and frontotemporal dementia. These perturbations in KIF1A transport, in tandem with Tau dysfunction, present compelling evidence of important relationship between these two proteins. To investigate and define the relationship between KIF1A and Tau, a single-molecule in vitro reconstituted system approach was employed. In doing so, an unexpected finding occurred in the early stages of experimentation. It was revealed that KIF1A exhibits a unique pausing behavior between segments of processive movement on the microtubule surface. This behavior, which had been unreported and uncharacterized, contradicts the canonical behavior of other well studied kinesin proteins. KIF1A pausing was found to be mediated by the C-terminal tail (CTT) structure of tubulin, the building blocks of microtubules. Further exploration revealed that KIF1A pausing was reliant upon the level of polyglutamylation, a post-translational modification enriched on neuronal tubulin, of the CTTs. Lastly, it was determined that polyglutamylated CTTs allow for an electrostatic tethering mechanism with the K-loop, a motor domain surface loop of KIF1A, that allows the motor to pause. Like KIF1A, Tau also relies on the tubulin CTTs to exhibit its characteristic diffusive binding behavior. In considering this fact, KIF1A regulation via Tau’s behavioral binding state was investigated. Ultimately, it was discovered that the diffusive binding state of Tau regulates KIF1A, not the static binding state. This regulation occurs when KIF1A tries to engage in a pause, but cannot due to diffusive Tau occupying the CTTs. This work provides a new mechanism of Tau-mediated kinesin motor regulation and the first direct link between KIF1A and Tau function, expanding our knowledge of the spatiotemporal regulation of axonal transport

A New Analysis Method for Reconstructing the Arrival Direction of TeV Gamma-rays Using a Single Imaging Atmospheric Cherenkov Telescope

We present a method of atmospheric Cherenkov imaging which reconstructs the unique arrival direction of TeV gamma rays using a single telescope. The method is derived empirically and utilizes several features of gamma-ray induced air showers which determine, to a precision of 0.12 degrees, the arrival direction of photons, on an event-by-event basis. Data from the Whipple Observatory's 10 m gamma-ray telescope is utilized to test selection methods based on source location. The results compare these selection methods with traditional techniques and three different camera fields of view. The method will be discussed in the context of a search for a gamma-ray signal from a point source located anywhere within the field of view and from regions of extended emission.Comment: 24 pages, 16 figures, accepted for publication in Astroparticle Physics May 11, 200

Analysis of Compression Pad Cavities for the Orion Heatshield

Current results of a program for analysis of the compression pad cavities on the Orion heatshield are reviewed. The program was supported by experimental tests, engineering modeling, and applied computations with an emphasis on the latter presented in this paper. The computational tools and approach are described along with calculated results for wind tunnel and flight conditions. Correlations of the computed results are shown which can produce a credible prediction of heating augmentation due to cavity disturbances. The models developed for use in preliminary design of the Orion heatshield are presented

A Superheated Droplet Detector for Dark Matter Search

We discuss the operation principle of a detector based on superheated droplets of Freon-12 and its feasibility for the search of weakly interacting cold dark matter particles. In particular we are interested in a neutralino search experiment in the mass range from 10 to 10^4 GeV/c^2 and with a sensitivity of better than 10^-2 events/kg/d. We show that our new proposed detector can be operated at ambient pressure and room temperature in a mode where it is exclusively sensitive to nuclear recoils like those following neutralino interactions, which allows a powerful background discrimination. An additional advantage of this technique is due to the fact that the detection material, Freon-12, is cheap and readily available in large quantities. Moreover we were able to show that piezoelectric transducers allow efficient event localization in large volumes.Comment: 15 pages LATEX; 11 figures on request from [email protected] submitted to Nuclear Instruments and Methods

Status of the PICASSO Project

The Picasso project is a dark matter search experiment based on the superheated droplet technique. Preliminary runs performed at the Picasso Lab in Montreal have showed the suitability of this detection technique to the search for weakly interacting cold dark matter particles. In July 2002, a new phase of the project started. A batch of six 1-liter detectors with an active mass of approximately 40g was installed in a gallery of the SNO observatory in Sudbury, Ontario, Canada at a depth of 6,800 feet (2,070m). We give a status report on the new experimental setup, data analysis, and preliminary limits on spin-dependent neutralino interaction cross section.Comment: 3 pages, 2 figures. To appear in the Proceedings of the TAUP 2003 conference, 5-9 September, 2003, University of Washington, Seattle, US