Global distributions of diazotrophs abundance, biomass and nitrogen fixation rates - Collection of source datasets - Contribution to the MAREDAT World Ocean Atlas of Plankton Functional Types

The MAREDAT atlas covers 11 types of plankton, ranging in size from bacteria to jellyfish. Together, these plankton groups determine the health and productivity of the global ocean and play a vital role in the global carbon cycle. Working within a uniform and consistent spatial and depth grid (map) of the global ocean, the researchers compiled thousands and tens of thousands of data points to identify regions of plankton abundance and scarcity as well as areas of data abundance and scarcity. At many of the grid points, the MAREDAT team accomplished the difficult conversion from abundance (numbers of organisms) to biomass (carbon mass of organisms). The MAREDAT atlas provides an unprecedented global data set for ecological and biochemical analysis and modeling as well as a clear mandate for compiling additional existing data and for focusing future data gathering efforts on key groups in key areas of the ocean. The present collection presents the original data sets used to compile Global distributions of diazotrophs abundance, biomass and nitrogen fixation rates

Global distributions of diazotrophs abundance and biomass - Depth integrated values computed from a collection of source datasets - Contribution to the MAREDAT World Ocean Atlas of Plankton Functional Types

The MAREDAT atlas covers 11 types of plankton, ranging in size from bacteria to jellyfish. Together, these plankton groups determine the health and productivity of the global ocean and play a vital role in the global carbon cycle. Working within a uniform and consistent spatial and depth grid (map) of the global ocean, the researchers compiled thousands and tens of thousands of data points to identify regions of plankton abundance and scarcity as well as areas of data abundance and scarcity. At many of the grid points, the MAREDAT team accomplished the difficult conversion from abundance (numbers of organisms) to biomass (carbon mass of organisms). The MAREDAT atlas provides an unprecedented global data set for ecological and biochemical analysis and modeling as well as a clear mandate for compiling additional existing data and for focusing future data gathering efforts on key groups in key areas of the ocean. The present data set presents depth integrated values of diazotrophs abundance and biomass, computed from a collection of source data sets.

Nitrogen fixation and nitrogen gene abundance of METEOR cruises M77/3 and M77/4

Nitrogen fixation, the biological reduction of dinitrogen gas (N2) to ammonium (NH4+), is quantitatively the most important external source of new nitrogen (N) to the open ocean. Classically, the ecological niche of oceanic N2 fixers (diazotrophs) is ascribed to tropical oligotrophic surface waters, often depleted in fixed N, with a diazotrophic community dominated by cyanobacteria. Although this applies for large areas of the ocean, biogeochemical models and phylogenetic studies suggest that the oceanic diazotrophic niche may be much broader than previously considered, resulting in major implications for the global N-budget. Here, we report on the composition, distribution and abundance of nifH, the functional gene marker for N2 fixation. Our results show the presence of eight clades of diazotrophs in the oxygen minimum zone (OMZ) off Peru. Although proteobacterial clades dominated overall, two clusters affiliated to spirochaeta and archaea were identified. N2 fixation was detected within OMZ waters and was stimulated by the addition of organic carbon sources supporting the view that non-phototrophic diazotrophs were actively fixing dinitrogen. The observed co-occurrence of key functional genes for N2 fixation, nitrification, anammox and denitrification suggests that a close spatial coupling of N-input and N-loss processes exists in the OMZ off Peru. The wide distribution of diazotrophs throughout the water column adds to the emerging view that the habitat of marine diazotrophs can be extended to low oxygen/high nitrate areas. Furthermore, our statistical analysis suggests that NO2- and PO43- are the major factors affecting diazotrophic distribution throughout the OMZ. In view of the predicted increase in ocean deoxygenation resulting from global warming, our findings indicate that the importance of OMZs as niches for N2 fixation may increase in the futur

Relative abundance of anaerobic methanotrophic archaea in bacterial mat Cascadia margin off Oregon

The microbially mediated anaerobic oxidation of methane (AOM) is the major biological sink of the greenhouse gas methane in marine sediments (doi:10.1007/978-94-009-0213-8_44) and serves as an important control for emission of methane into the hydrosphere. The AOM metabolic process is assumed to be a reversal of methanogenesis coupled to the reduction of sulfate to sulfide involving methanotrophic archaea (ANME) and sulfate-reducing bacteria (SRB) as syntrophic partners which were describes amongst others in Boetius et al. (2000; doi:10.1038/35036572). In this study, 16S rRNA-based methods were used to investigate the distribution and biomass of archaea in samples from sediments above outcropping methane hydrate at Hydrate Ridge (Cascadia margin off Oregon) and (ii) massive microbial mats enclosing carbonate reefs (Crimea area, Black Sea). Sediment samples from Hydrate Ridge were obtained during R/V SONNE cruises SO143-2 in August 1999 and SO148-1 in August 2000 at the crest of southern Hydrate Ridge at the Cascadia convergent margin off the coast of Oregon. The second study area is located in the Black Sea and represents a field in which there is active seepage of free gas on the slope of the northwestern Crimea area. Here, a field of conspicuous microbial reefs forming chimney-like structures was discovered at a water depth of 230 m in anoxic waters. The microbial mats were sampled by using the manned submersible JAGO during the R/V Prof. LOGACHEV cruise in July 2001. At Hydrate Ridge the surface sediments were dominated by aggregates consisting of ANME-2 and members of the Desulfosarcina-Desulfococcus branch (DSS) (ANME-2/DSS aggregates), which accounted for >90% of the total cell biomass. The numbers of ANME-1 cells increased strongly with depth; these cells accounted 1% of all single cells at the surface and more than 30% of all single cells (5% of the total cells) in 7- to 10-cm sediment horizons that were directly above layers of gas hydrate. In the Black Sea microbial mats ANME-1 accounted for about 50% of all cells. ANME-2/DSS aggregates occurred in microenvironments within the mat but accounted for only 1% of the total cells. FISH probes for the ANME-2a and ANME-2c subclusters were designed based on a comparative 16S rRNA analysis. In Hydrate Ridge sediments ANME-2a/DSS and ANME-2c/DSS aggregates differed significantly in morphology and abundance. The relative abundance values for these subgroups were remarkably different at Beggiatoa sites (80% ANME-2a, 20% ANME-2c) and Calyptogena sites (20% ANME-2a, 80% ANME-2c), indicating that there was preferential selection of the groups in the two habitats.

(Table S2) Relative abundance of archaeal 16S rRNA genes in sediment cores

Deep drilling into the marine sea floor has uncovered a vast sedimentary ecosystem of microbial cells (Parkes et al., 1994, doi:10.1038/371410a0; D'Hondt et al., 2004, doi:10.1126/science.1101155). Extrapolation of direct counts of stained microbial cells to the total volume of habitable marine subsurface sediments suggests that between 56 Pg (Parkes et al., 1994, doi:10.1038/371410a0) and 303 Pg (Whitman et al., 1998) of cellular carbon could be stored in this largely unexplored habitat. From recent studies using various culture-independent techniques, no clear picture has yet emerged as to whether Archaea or Bacteria are more abundant in this extensive ecosystem (Schippers et al., doi:10.1038/nature03302; Inagaki et al., doi:10.1073/pnas.0511033103 ; Mauclaire et al., doi:10.1111/j.1472-4677.2004.00035.x; Biddle et al., doi:10.1073/pnas.0600035103). Here we show that in subsurface sediments buried deeper than 1 m in a wide range of oceanographic settings at least 87% of intact polar membrane lipids, biomarkers for the presence of live cells (Biddle et al., doi:10.1073/pnas.0600035103; Sturt et al., 2004, doi:10.1002/rcm.1378), are attributable to archaeal membranes, suggesting that Archaea constitute a major fraction of the biomass. Results obtained from modified quantitative polymerase chain reaction and slot-blot hybridization protocols support the lipid-based evidence and indicate that these techniques have previously underestimated archaeal biomass. The lipid concentrations are proportional to those of total organic carbon. On the basis of this relationship, we derived an independent estimate of amounts of cellular carbon in the global marine subsurface biosphere. Our estimate of 90 Pg of cellular carbon is consistent, within an order of magnitude, with previous estimates, and underscores the importance of marine subsurface habitats for global biomass budgets.

Relative abundance of diatom species in surface samples from the Southern Ocean, Sea ice species

Diatoms from 228 Southern Ocean core-top sediment samples were examined to determine the geographic distributions of 32 major diatom species/taxa preserved in the sediments of three zonally-distinct regions; Sea Ice, Open Ocean and the Tropical/Subtropical. In the first of three papers, 14 species/taxa occurring in the region where sea ice covers the ocean surface on an annual basis are geographically documented. Comparisons are drawn between the diatom abundances on the sea floor, sea ice parameters (annual duration and concentration in February and September) and February sea-surface temperature. Such parameters are commonly used in reconstructing past oceanographic conditions in the Sea Ice and Open Ocean zones. Analysis of the geographic patterns and sea-surface parameter correlations reveals species-specific distributions regulated primarily by sea ice coverage and sea-surface temperature, which support the use of diatom remains for the estimation of these past sea-surface environmental parameters. Comparison with reliable accounts of the 14 species from the sediments or plankton also provides the first glimpses into species-specific ecology and habitat linkages.

Relative abundance of euphausiids (Crustacea) in water samples of the Arabian Sea (Table 2)

In the present paper, the ecology and feeding habits of euphausiids are described. The samples were taken at the time of the NE-monsoon (1964/65) by R. V. "Meteor" in the Arabian Sea and adjacent waters. 24 species were determined. According to distribution of the species, the following marine areas can be distinguished: Arabian Sea: 24 species, dominant are Euphausia diomedeae, E. tenera, E. distinguenda, Stylocheiron carinatum. Gulf of Aden: 10 species, dominant are Euphausia diomedeae, E. distinguenda. Red Sea: 6 species, dominant are Euphausia diomedeae, E. distinguenda. Gulf of Oman : 5 Species, dominant are Euphausia distinguenda, Pseudeupbaufia latifrons. Persian Gulf: 1 species - Pseudeuphausia latifrons. The total number of euphausiids indicate the biomass of this group. High densities of euphausiids (200-299 and > 300 individuals/100 m**3) occur in the innermost part of the Gulf cf Aden, in the area south of the equator near the African east coast, near Karachi (Indian west coast) and in the Persian Gulf. Comparison with data relating to production biology confirms that these are eutrophic zones which coincide with areas in which upwelling occurs at the time of the NE-monsoon. The central part of the Arabian Sea differs from adjacent waters by virtue of less dense euphausiid populations (> 199 individuals/100 m**3). Measurements relating to production biology demonstrate a relatively low concentration of primary food sources. Food material was ascertained by analysis of stomach content. The following omnivorous species were examined: Euphausia diomedeae, E. distinguenda, E. tenera, Pseudeuphausia latifrons and Thysanopoda tricuspidata. Apart from crustacean remains large numbers of Foraminifera, Radiolaria, tintinnids, dinoflagellates were found in the stomachs. Quantitatively crustaceans form the most important item in the diet. Food selection on the basis of size and form appears to be restricted to certain genera of tintinnids. The genera Stylocheiron and Nematoscelis are predators. Only crustacean remains were found in the stomachs of Stylocheiron abbreviatum, whereas Radiolaria, Foraminifera and tintinnids occurred to some extent in Nematasceli sp. Different euphausiids in the food chain in the Arabian Sea. In omnivorous species the position is variable, since they not only feed by filtering autotrophic and heterotrophic Protista, but also by predation on zooplankton. Carnivorous species without filtering apparatus feed exclusively on zooplankton of the size of copepods. Only these species are well established as occupying a higher position in the food chain. The parasitic protozoan Tbalassomyces fagei was found on Euphausia diomedeae, E. fenera, E. distinguenda and E. sanzoi.

(Appendix A) Radiolarian abundance in ODP Hole 202-1241A

A quantitative radiolarian study at Ocean Drilling Program Site 1241 in the eastern tropical Pacific enables us to reconstruct paleoceanographic changes that occurred since the latest middle Miocene. Today, this site is located just under the Eastern Pacific Warm Pool (EPWP). Based on the abundance variations of radiolarian characteristic species which are indicators of upwelling and thermocline changes, it is suggested that three notable changes occurred at 10.6, 9.8, and 4.2 Ma in the region. Four distinct periods of oceanographic conditions bounded by these notable changes were characterized on the basis of the following: (1) stratified seawater (12.0 to 10.6 Ma); (2) a shallowing of the thermocline and an increasing of upwelling (10.6 to 9.8 Ma); (3) significant inflow of warm water to the eastern tropical Pacific caused by an intensified Northern Equatorial Countercurrent (NECC), resulting in the formation of EPWP (9.8 to 4.2 Ma); and (4) the reduction of the EPWP and the NECC, and an increase in upwelling (4.2 to 0 Ma). The timing of these paleoceanographic events indicated the strong relations with the opening and closing of the Indonesian and Central American (Panama) Seaways. The reduction of the EPWP (this study) and the deepening of the thermocline in western Pacific at about 4.2 Ma (Cannariato and Ravelo, 1997; Chaisson and Ravelo, 2000) indicated a change from a state resembling El Niño in the late Miocene and the early Pliocene time to a state resembling La Niña by the late Pliocene

Global distributions of diazotrophs abundance, biomass and nitrogen fixation rates - Gridded data product (NetCDF) - Contribution to the MAREDAT World Ocean Atlas of Plankton Functional Types

The MAREDAT atlas covers 11 types of plankton, ranging in size from bacteria to jellyfish. Together, these plankton groups determine the health and productivity of the global ocean and play a vital role in the global carbon cycle. Working within a uniform and consistent spatial and depth grid (map) of the global ocean, the researchers compiled thousands and tens of thousands of data points to identify regions of plankton abundance and scarcity as well as areas of data abundance and scarcity. At many of the grid points, the MAREDAT team accomplished the difficult conversion from abundance (numbers of organisms) to biomass (carbon mass of organisms). The MAREDAT atlas provides an unprecedented global data set for ecological and biochemical analysis and modeling as well as a clear mandate for compiling additional existing data and for focusing future data gathering efforts on key groups in key areas of the ocean. This is a gridded data product about diazotrophic organisms . There are 6 variables. Each variable is gridded on a dimension of 360 (longitude) * 180 (latitude) * 33 (depth) * 12 (month). The first group of 3 variables are: (1) number of biomass observations, (2) biomass, and (3) special nifH-gene-based biomass. The second group of 3 variables is same as the first group except that it only grids non-zero data. We have constructed a database on diazotrophic organisms in the global pelagic upper ocean by compiling more than 11,000 direct field measurements including 3 sub-databases: (1) nitrogen fixation rates, (2) cyanobacterial diazotroph abundances from cell counts and (3) cyanobacterial diazotroph abundances from qPCR assays targeting nifH genes. Biomass conversion factors are estimated based on cell sizes to convert abundance data to diazotrophic biomass. Data are assigned to 3 groups including Trichodesmium, unicellular diazotrophic cyanobacteria (group A, B and C when applicable) and heterocystous cyanobacteria (Richelia and Calothrix). Total nitrogen fixation rates and diazotrophic biomass are calculated by summing the values from all the groups. Some of nitrogen fixation rates are whole seawater measurements and are used as total nitrogen fixation rates. Both volumetric and depth-integrated values were reported. Depth-integrated values are also calculated for those vertical profiles with values at 3 or more depths.

Are critically endangered fish back on the menu? Analysis of U.K. fisheries data suggest post-ban landings of prohibited skates in European waters

Skates (Rajidae) have been commercially exploited in Europe for hundreds of years with some species’ abundances declining dramatically during the twentieth century. In 2009 it became “prohibited for EU vessels to target, retain, tranship or land” certain species in some ICES areas, including the critically endangered common skate and the endangered white skate. To examine compliance with skate bans the official UK landings data for 2011–2014 were analysed. Surprisingly, it was found that after the ban prohibited species were still reported landed in UK ports, including 9.6 t of common skate during 2011–2014. The majority of reported landings of common and white skate were from northern UK waters and landed into northern UK ports. Although past landings could not be validated as being actual prohibited species, the landings’ patterns found reflect known abundance distributions that suggest actual landings were made, rather than sporadic occurrence across ports that would be evident if landings were solely due to systematic misidentification or data entry errors. Nevertheless, misreporting and data entry errors could not be discounted as factors contributing to the recorded landings of prohibited species. These findings raise questions about the efficacy of current systems to police skate landings to ensure prohibited species remain protected. By identifying UK ports with the highest apparent landings of prohibited species and those still landing species grouped as'skates and rays’, these results may aid authorities in allocating limited resources more effectively to reduce landings, misreporting and data errors of prohibited species, and increase species-specific landing compliance.

Are critically endangered fish back on the menu? Analysis of U.K. fisheries data suggest post-ban landings of prohibited skates in European waters

Skates (Rajidae) have been commercially exploited in Europe for hundreds of years with some species’ abundances declining dramatically during the twentieth century. In 2009 it became “prohibited for EU vessels to target, retain, tranship or land” certain species in some ICES areas, including the critically endangered common skate and the endangered white skate. To examine compliance with skate bans the official UK landings data for 2011–2014 were analysed. Surprisingly, it was found that after the ban prohibited species were still reported landed in UK ports, including 9.6 t of common skate during 2011–2014. The majority of reported landings of common and white skate were from northern UK waters and landed into northern UK ports. Although past landings could not be validated as being actual prohibited species, the landings’ patterns found reflect known abundance distributions that suggest actual landings were made, rather than sporadic occurrence across ports that would be evident if landings were solely due to systematic misidentification or data entry errors. Nevertheless, misreporting and data entry errors could not be discounted as factors contributing to the recorded landings of prohibited species. These findings raise questions about the efficacy of current systems to police skate landings to ensure prohibited species remain protected. By identifying UK ports with the highest apparent landings of prohibited species and those still landing species grouped as'skates and rays’, these results may aid authorities in allocating limited resources more effectively to reduce landings, misreporting and data errors of prohibited species, and increase species-specific landing compliance.

Development of Imaging Mass Spectrometry Analysis of Lipids in Biological and Clinically Relevant Applications

La spectrométrie de masse mesure la masse des ions selon leur rapport masse sur charge. Cette technique est employée dans plusieurs domaines et peut analyser des mélanges complexes. L’imagerie par spectrométrie de masse (Imaging Mass Spectrometry en anglais, IMS), une branche de la spectrométrie de masse, permet l’analyse des ions sur une surface, tout en conservant l’organisation spatiale des ions détectés. Jusqu’à présent, les échantillons les plus étudiés en IMS sont des sections tissulaires végétales ou animales. Parmi les molécules couramment analysées par l’IMS, les lipides ont suscité beaucoup d'intérêt. Les lipides sont impliqués dans les maladies et le fonctionnement normal des cellules; ils forment la membrane cellulaire et ont plusieurs rôles, comme celui de réguler des événements cellulaires. Considérant l’implication des lipides dans la biologie et la capacité du MALDI IMS à les analyser, nous avons développé des stratégies analytiques pour la manipulation des échantillons et l’analyse de larges ensembles de données lipidiques. La dégradation des lipides est très importante dans l’industrie alimentaire. De la même façon, les lipides des sections tissulaires risquent de se dégrader. Leurs produits de dégradation peuvent donc introduire des artefacts dans l’analyse IMS ainsi que la perte d’espèces lipidiques pouvant nuire à la précision des mesures d’abondance. Puisque les lipides oxydés sont aussi des médiateurs importants dans le développement de plusieurs maladies, leur réelle préservation devient donc critique. Dans les études multi-institutionnelles où les échantillons sont souvent transportés d’un emplacement à l’autre, des protocoles adaptés et validés, et des mesures de dégradation sont nécessaires. Nos principaux résultats sont les suivants : un accroissement en fonction du temps des phospholipides oxydés et des lysophospholipides dans des conditions ambiantes, une diminution de la présence des lipides ayant des acides gras insaturés et un effet inhibitoire sur ses phénomènes de la conservation des sections au froid sous N2. A température et atmosphère ambiantes, les phospholipides sont oxydés sur une échelle de temps typique d’une préparation IMS normale (~30 minutes). Les phospholipides sont aussi décomposés en lysophospholipides sur une échelle de temps de plusieurs jours. La validation d’une méthode de manipulation d’échantillon est d’autant plus importante lorsqu’il s’agit d’analyser un plus grand nombre d’échantillons. L’athérosclérose est une maladie cardiovasculaire induite par l’accumulation de matériel cellulaire sur la paroi artérielle. Puisque l’athérosclérose est un phénomène en trois dimension (3D), l'IMS 3D en série devient donc utile, d'une part, car elle a la capacité à localiser les molécules sur la longueur totale d’une plaque athéromateuse et, d'autre part, car elle peut identifier des mécanismes moléculaires du développement ou de la rupture des plaques. l'IMS 3D en série fait face à certains défis spécifiques, dont beaucoup se rapportent simplement à la reconstruction en 3D et à l’interprétation de la reconstruction moléculaire en temps réel. En tenant compte de ces objectifs et en utilisant l’IMS des lipides pour l’étude des plaques d’athérosclérose d’une carotide humaine et d’un modèle murin d’athérosclérose, nous avons élaboré des méthodes «open-source» pour la reconstruction des données de l’IMS en 3D. Notre méthodologie fournit un moyen d’obtenir des visualisations de haute qualité et démontre une stratégie pour l’interprétation rapide des données de l’IMS 3D par la segmentation multivariée. L’analyse d’aortes d’un modèle murin a été le point de départ pour le développement des méthodes car ce sont des échantillons mieux contrôlés. En corrélant les données acquises en mode d’ionisation positive et négative, l’IMS en 3D a permis de démontrer une accumulation des phospholipides dans les sinus aortiques. De plus, l’IMS par AgLDI a mis en évidence une localisation différentielle des acides gras libres, du cholestérol, des esters du cholestérol et des triglycérides. La segmentation multivariée des signaux lipidiques suite à l’analyse par IMS d’une carotide humaine démontre une histologie moléculaire corrélée avec le degré de sténose de l’artère. Ces recherches aident à mieux comprendre la complexité biologique de l’athérosclérose et peuvent possiblement prédire le développement de certains cas cliniques. La métastase au foie du cancer colorectal (Colorectal cancer liver metastasis en anglais, CRCLM) est la maladie métastatique du cancer colorectal primaire, un des cancers le plus fréquent au monde. L’évaluation et le pronostic des tumeurs CRCLM sont effectués avec l’histopathologie avec une marge d’erreur. Nous avons utilisé l’IMS des lipides pour identifier les compartiments histologiques du CRCLM et extraire leurs signatures lipidiques. En exploitant ces signatures moléculaires, nous avons pu déterminer un score histopathologique quantitatif et objectif et qui corrèle avec le pronostic. De plus, par la dissection des signatures lipidiques, nous avons identifié des espèces lipidiques individuelles qui sont discriminants des différentes histologies du CRCLM et qui peuvent potentiellement être utilisées comme des biomarqueurs pour la détermination de la réponse à la thérapie. Plus spécifiquement, nous avons trouvé une série de plasmalogènes et sphingolipides qui permettent de distinguer deux différents types de nécrose (infarct-like necrosis et usual necrosis en anglais, ILN et UN, respectivement). L’ILN est associé avec la réponse aux traitements chimiothérapiques, alors que l’UN est associé au fonctionnement normal de la tumeur.

No Joke: Understanding Public Sentiment Toward Selling and Salespeople Through Cartoon Analysis

Unflattering representations of salesmanship in mass media exist in abundance. In order to gauge the depiction of selling in mass media, this article explores the nature and public perceptions of salesmanship using editorial cartoons. A theory of cartooning suggests that editorial cartoons reflect public sentiment toward events and issues and therefore provide a useful way of measuring and tracking such sentiment over time. The criteria of narrative, location, binary struggle, normative transference, and metaphor were used as a framework to analyze 286 cartoons over a 30-year period from 1983 to 2013. The results suggest that while representations of the characteristics and behaviors of salespeople shifted very little across time periods, changes in public perceptions of seller–buyer conflict, the role of the customer, and selling techniques were observed, thus indicating that cartoons are sensitive enough to measure the portrayal of selling.

On independent component analysis applied to unmixing hyperspectral data

One of the most challenging task underlying many hyperspectral imagery applications is the spectral unmixing, which decomposes a mixed pixel into a collection of reectance spectra, called endmember signatures, and their corresponding fractional abundances. Independent Component Analysis (ICA) have recently been proposed as a tool to unmix hyperspectral data. The basic goal of ICA is to nd a linear transformation to recover independent sources (abundance fractions) given only sensor observations that are unknown linear mixtures of the unobserved independent sources. In hyperspectral imagery the sum of abundance fractions associated to each pixel is constant due to physical constraints in the data acquisition process. Thus, sources cannot be independent. This paper address hyperspectral data source dependence and its impact on ICA performance. The study consider simulated and real data. In simulated scenarios hyperspectral observations are described by a generative model that takes into account the degradation mechanisms normally found in hyperspectral applications. We conclude that ICA does not unmix correctly all sources. This conclusion is based on the a study of the mutual information. Nevertheless, some sources might be well separated mainly if the number of sources is large and the signal-to-noise ratio (SNR) is high.