Increased bacterial growth efficiency with environmental variability: results from DOC degradation by bacteria in pure culture experiments.

This paper assesses how considering variation in DOC availability and cell maintenance in bacterial models affects Bacterial Growth Efficiency (BGE) estimations. For this purpose, we conducted two biodegradation experiments simultaneously. In experiment one, a given amount of substrate was added to the culture at the start of the experiment whilst in experiment two, the same amount of substrate was added, but using periodic pulses over the time course of the experiment. Three bacterial models, with different levels of complexity, (the Monod, Marr-Pirt and the dynamic energy budget – DEB – models), were used and calibrated using the above experiments. BGE has been estimated using the experimental values obtained from discrete samples and from model generated data. Cell maintenance was derived experimentally, from respiration rate measurements. The results showed that the Monod model did not reproduce the experimental data accurately, whereas the Marr-Pirt and DEB models demonstrated a good level of reproducibility, probably because cell maintenance was built into their formula. Whatever estimation method was used, the BGE value was always higher in experiment two (the periodically pulsed substrate) as compared to the initially one-pulsed-substrate experiment. Moreover, BGE values estimated without considering cell maintenance (Monod model and empirical formula) were always smaller than BGE values obtained from models taking cell maintenance into account. Since BGE is commonly estimated using constant experimental systems and ignore maintenance, we conclude that these typical methods underestimate BGE values. On a larger scale, and for biogeochemical cycles, this would lead to the conclusion that, for a given DOC supply rate and a given DOC consumption rate, these BGE estimation methods overestimate the role of bacterioplankton as CO<sub>2</sub> producers

Diffraction microtomography with sample rotation: influence of a missing apple core in the recorded frequency space

Diffraction microtomography in coherent light is foreseen as a promising technique to image transparent living samples in three dimensions without staining. Contrary to conventional microscopy with incoherent light, which gives morphological information only, diffraction microtomography makes it possible to obtain the complex optical refractive index of the observed sample by mapping a three-dimensional support in the spatial frequency domain. The technique can be implemented in two configurations, namely, by varying the sample illumination with a fixed sample or by rotating the sample using a fixed illumination. In the literature, only the former method was described in detail. In this report, we precisely derive the three-dimensional frequency support that can be mapped by the sample rotation configuration. We found that, within the first-order Born approximation, the volume of the frequency domain that can be mapped exhibits a missing part, the shape of which resembles that of an apple core. The projection of the diffracted waves in the frequency space onto the set of sphere caps covered by the sample rotation does not allow for a complete mapping of the frequency along the axis of rotation due to the finite radius of the sphere caps. We present simulations of the effects of this missing information on the reconstruction of ideal objects.Comment: 7 pages, 11 figures, presented at Focus On Microscopy 200

Human skin in vitro permeation of bentazon and isoproturon formulations with or without protective clothing suit

Skin exposures to chemicals may lead, through percutaneous permeation, to a significant increase in systemic circulation. Skin is the primary route of entry during some occupational activities, especially in agriculture. To reduce skin exposures, the use of personal protective equipment (PPE) is recommended. PPE efficiency is characterized as the time until products permeate through material (lag time, Tlag). Both skin and PPE permeations are assessed using similar in vitro methods; the diffusion cell system. Flow-through diffusion cells were used in this study to assess the permeation of two herbicides, bentazon and isoproturon, as well as four related commercial formulations (Basagran(®), Basamais(®), Arelon(®) and Matara(®)). Permeation was measured through fresh excised human skin, protective clothing suits (suits) (Microchem(®) 3000, AgriSafe Pro(®), Proshield(®) and Microgard(®) 2000 Plus Green), and a combination of skin and suits. Both herbicides, tested by itself or as an active ingredient in formulations, permeated readily through human skin and tested suits (Tlag < 2 h). High permeation coefficients were obtained regardless of formulations or tested membranes, except for Microchem(®) 3000. Short Tlag, were observed even when skin was covered with suits, except for Microchem(®) 3000. Kp values tended to decrease when suits covered the skin (except when Arelon(®) was applied to skin covered with AgriSafe Pro and Microgard(®) 2000), suggesting that Tlag alone is insufficient in characterizing suits. To better estimate human skin permeations, in vitro experiments should not only use human skin but also consider the intended use of the suit, i.e., the active ingredient concentrations and type of formulations, which significantly affect skin permeation

Paenibacillus melissococcoides sp. nov., isolated from a honey bee colony affected by European foulbrood disease.

A novel, facultatively anaerobic, Gram-stain-positive, motile, endospore-forming bacterium of the genus Paenibacillus, designated strain 2.1T, was isolated from a colony of Apis mellifera affected by European foulbrood disease in Switzerland. The rod-shaped cells of strain 2.1T were 2.2–6.5 μm long and 0.7–1.1 μm wide. Colonies of strain 2.1T were orange-pigmented under oxic growth conditions on solid basal medium at 35–37 °C. Strain 2.1T showed catalase and cytochrome c oxidase activity. Its polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, aminophospholipid and phospholipid. The only respiratory quinone was menaquinone 7, and the major cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0, iso-C15 : 0, iso-C17 : 0 and palmitic acid (C16 : 0), which is consistent with other members of the genus Paenibacillus. The G+C content of the genomic DNA of strain 2.1T was 53.3 mol%. Phylogenetic analyses based on the 16S rRNA gene sequence similarity showed that strain 2.1T was closely related to Paenibacillus dendritiformis LMG 21716T (99.7 % similarity) and Paenibacillus thiaminolyticus DSM 7262T (98.8 %). The whole-genome average nucleotide identity between strain 2.1T and the type strains of P. dendritiformis and P. thiaminolyticus was 92 and 91 %, respectively, and thus lower than the 95 % threshold value for delineation of genomic prokaryotic species. Based on the results of phylogenetic, genomic, phenotypic and chemotaxonomic analyses we propose the name Paenibacillus melissococcoides sp. nov. for this novel Paenibacillus species. The type strain is 2.1T (=CCOS 2000T=DSM 113619T=LMG 32539T)

Dinitrogen fixation and dissolved organic nitrogen fueled primary production and particulate export during the VAHINE mesocosm experiment (New Caledonia lagoon)

International audienceIn the oligotrophic ocean characterized by nitrate (NO − 3) depletion in surface waters, dinitrogen (N 2) fixation and dissolved organic nitrogen (DON) can represent significant nitrogen (N) sources for the ecosystem. In this study, we deployed large in situ mesocosms in New Caledonia in order to investigate (1) the contribution of N 2 fixation and DON use to primary production (PP) and particle export and (2) the fate of the freshly produced particulate organic N (PON), i.e., whether it is preferentially accumulated and recycled in the water column or exported out of the system. The mesocosms were fertilized with phosphate (PO 3− 4) in order to prevent phosphorus (P) limitation and promote N 2 fixation. The diazotrophic community was dominated by diatom–diazotroph associations (DDAs) during the first part of the experiment for 10 days (P1) followed by the unicel-lular N 2-fixing cyanobacteria UCYN-C for the last 9 days (P2) of the experiment. N 2 fixation rates averaged 9.8 ± 4.0 and 27.7 ± 8.6 nmol L −1 d −1 during P1 and P2, respectively. NO − 3 concentrations ( 0.05) during P1 (9.0 ± 3.3 %) and P2 (12.6 ± 6.1 %). However, the e ratio that quantifies the efficiency of a system to export particulate organic carbon (POC export) compared to PP (e ratio = POC export / PP) was significantly higher (p 0.05) from the total amount of PON exported (0.10 ± 0.04 µmol L −1), suggesting a rapid and probably direct export of the recently fixed N 2 by the DDAs. During P2, both PON concentrations and PON export increased in the mesocosms by a factor 1.5–2. Unlike in P1, this PON production was not totally explained by the new N provided by N 2 fixation. The use of DON, whose concentrations decreased significantly (p < 0.05) from 5.3 ± 0.5 µmol L −1 to 4.4 ± 0.5 µmol L −1 , appeared to be the missing N source. The DON consumption (∼ 0.9 µmol L −1) during P2 is higher Published by Copernicus Publications on behalf of the European Geosciences Union. 4100 H. Berthelot et al.: Dinitrogen fixation and dissolved organic nitrogen fueled primary production than the total amount of new N brought by N 2 fixation (∼ 0.25 µmol L −1) during the same period. These results suggest that while DDAs mainly rely on N 2 fixation for their N requirements, both N 2 fixation and DON can be significant N sources for primary production and particulate export following UCYN-C blooms in the New Caledonia lagoon and by extension in the N-limited oceans where similar events are likely to occur

Seasonal survey of the composition and degradation state of particulate organic matter in the Rhône River using lipid tracers

International audienceLipid tracers including fatty acids, hydroxyacids, n-alkanols, sterols and triterpenoids were used to determine the origin and fate of suspended particulate organic matter (POM) collected in the Rhône River (France). This seasonal survey (April 2011 to May 2013) revealed a year-round strong terrestrial higher-plant contribution to the particulate organic matter (POM), with significant algal inputs observed in March and attributed to phytoplanktonic blooms likely dominated by diatoms. Major terrigenous contributors to our samples are gymnosperms, and more precisely their roots and stems, as evidenced by the presence of high proportions of ω-hydroxydocosanoic acid (a suberin biomarker). The high amounts of coprostanol detected clearly show that the Rhône River is significantly affected by sewage waters. Specific sterol degradation products were quantified and used to assess the part of biotic and abiotic degradation of POM within the river. Higher-plant-derived organic matter appears to be mainly affected by photo-oxidation and au-toxidation (free radical oxidation), while organic matter of mammal or human origin, evidenced by the presence of co-prostanol, is clearly more prone to bacterial degradation. Despite the involvement of an intense autoxidation-inducing ho-molytic cleavage of peroxy bonds, a significant proportion of hydroperoxides is still intact in higher plant debris. These compounds could affect the degradation of terrestrial material by inducing an intense autoxidation upon its arrival at sea

PAI-1 and functional blockade of SNAI1 in breast cancer cell migration

12 pages, 5 figures.-- PMID: 19055748 [PubMed].-- et al.[Introduction]: Snail, a family of transcriptional repressors implicated in cell movement, has been correlated with tumour invasion. The Plasminogen Activation (PA) system, including urokinase plasminogen activator (uPA), its receptor and its inhibitor, plasminogen activator inhibitor type 1(PAI-1), also plays a key role in cancer invasion and metastasis, either through proteolytic degradation or by non-proteolytic modulation of cell adhesion and migration. Thus, Snail and the PA system are both over-expressed in cancer and influence this process. In this study we aimed to determine if the activity of SNAI1 (a member of the Snail family) is correlated with expression of the PA system components and how this correlation can influence tumoural cell migration.[Methods]: We compared the invasive breast cancer cell-line MDA-MB-231 expressing SNAI1 (MDA-mock) with its derived clone expressing a dominant-negative form of SNAI1 (SNAI1-DN). Expression of PA system mRNAs was analysed by cDNA microarrays and real-time quantitative RT-PCR. Wound healing assays were used to determine cell migration. PAI-1 distribution was assessed by immunostaining.[Results]: We demonstrated by both cDNA microarrays and realtime quantitative RT-PCR that the functional blockade of SNAI1 induces a significant decrease of PAI-1 and uPA transcripts. After performing an in vitro wound-healing assay, we observed that SNAI1-DN cells migrate more slowly than MDA-mock cells and in a more collective manner. The blockade of SNAI1 activity resulted in the redistribution of PAI-1 in SNAI1-DN cells decorating large lamellipodia, which are commonly found structures in these cells.[Conclusions]: In the absence of functional SNAI1, the expression of PAI-1 transcripts is decreased, although the protein is redistributed at the leading edge of migrating cells in a manner comparable with that seen in normal epithelial cells.This work was supported by the CNRS ACI Program "Complexité du vivant" (grant # 050009DR11) and by the Evry Genopole grant "Aide à l'acquisition d'équipement semi-lourd" 2007 and 2008.Peer reviewe

New live screening of plant-nematode interactions in the rhizosphere

Abstract Free living nematodes (FLN) are microscopic worms found in all soils. While many FLN species are beneficial to crops, some species cause significant damage by feeding on roots and vectoring viruses. With the planned legislative removal of traditionally used chemical treatments, identification of new ways to manage FLN populations has become a high priority. For this, more powerful screening systems are required to rapidly assess threats to crops and identify treatments efficiently. Here, we have developed new live assays for testing nematode responses to treatment by combining transparent soil microcosms, a new light sheet imaging technique termed Biospeckle Selective Plane Illumination Microscopy (BSPIM) for fast nematode detection, and Confocal Laser Scanning Microscopy for high resolution imaging. We show that BSPIM increased signal to noise ratios by up to 60 fold and allowed the automatic detection of FLN in transparent soil samples of 1.5 mL. Growing plant root systems were rapidly scanned for nematode abundance and activity, and FLN feeding behaviour and responses to chemical compounds observed in soil-like conditions. This approach could be used for direct monitoring of FLN activity either to develop new compounds that target economically damaging herbivorous nematodes or ensuring that beneficial species are not negatively impacted