A TGA/FT-IR study for OC and EC quantification applied to carbonaceous aerosol collected in Milan (Italy)

International audienceCarbon analysis consists in the evaluation of the carbonaceous content of the aerosol (TC) but, more importantly, of its distribution between the two components EC (Elemental Carbon) and OC (Organic Carbon) that are characterized by different physical-chemical properties. In spite of the numerous studies focused on this topic, nowadays, a universal methodology for the determination of the two components EC and OC is not available. In fact OC and EC (also known as black carbon or soot) are operationally defined by the method of analysis and, as a consequence, different methods can produce different results. In this paper we present results on the application of TGA/FT-IR (Thermogravimetric Analysis/Fourier Transformed Infrared Spectroscopy) to the characterization of carbonaceous aerosols. The analytical methodology was applied to PM10 four-hour time resolution samples collected in Milan urban area. The method is a two-steps thermal one and bases itself on the different thermal behaviour of OC and EC. It has been set up analyzing suitable standards containing both organic and elemental carbon. Carbon quantification is achieved by on-line, continuous monitoring of CO2 infrared absorption at 2361 cm?1. A good separation between OC and EC on particulate matter samples has been obtained. Ranges and average values were respectively 12?70 µg/m3 and 20 µg/m3 for OC and 0.2?6 µg/m3 and 2 µg/m3 for EC. On average OC and EC made up respectively 29.3 (±12.8) % and 2.5 (±1.8) % of PM10 fraction. The method reliability has been verified by comparison with TOT (Thermal Optical Transmission) technique. OC and EC values determined for ambient samples of PM10 were also correlated with meteorological parameters as well as with Radon concentrations

Inferring Foraging Areas of Nesting Loggerhead Turtles Using Satellite Telemetry and Stable Isotopes

In recent years, the use of intrinsic markers such as stable isotopes to link breeding and foraging grounds of migratory species has increased. Nevertheless, several assumptions still must be tested to interpret isotopic patterns found in the marine realm. We used a combination of satellite telemetry and stable isotope analysis to (i) identify key foraging grounds used by female loggerheads nesting in Florida and (ii) examine the relationship between stable isotope ratios and post-nesting migration destinations. We collected tissue samples for stable isotope analysis from 14 females equipped with satellite tags and an additional 57 untracked nesting females. Telemetry identified three post-nesting migratory pathways and associated non-breeding foraging grounds: (1) a seasonal continental shelf-constrained migratory pattern along the northeast U. S. coastline, (2) a non-breeding residency in southern foraging areas and (3) a residency in the waters adjacent to the breeding area. Isotopic variability in both delta C-13 and delta N-15 among individuals allowed identification of three distinct foraging aggregations. We used discriminant function analysis to examine how well delta C-13 and delta N-15 predict female post-nesting migration destination. The discriminant analysis classified correctly the foraging ground used for all but one individual and was used to predict putative feeding areas of untracked turtles. We provide the first documentation that the continental shelf of the Mid-and South Atlantic Bights are prime foraging areas for a large number (61%) of adult female loggerheads from the largest loggerhead nesting population in the western hemisphere and the second largest in the world. Our findings offer insights for future management efforts and suggest that this technique can be used to infer foraging strategies and residence areas in lieu of more expensive satellite telemetry, enabling sample sizes that are more representative at the population level

Enzymatic scarification of Anacamptis morio (Orchidaceae) seed facilitates lignin degradation, water uptake and germination

The seed coat of many species contains hydrophobic lignins, and in soil the action of microbial ligninases may contribute to release from dormancy. Laboratory use of ligninases to stimulate germination is promising because of the specific action on the seed coat, whereas chemical scarification agents may also corrode the embryo. We hypothesised that exposure of Anacamptis morio (Orchidaceae) seeds to fungal laccase would stimulate germination, and that the mechanism involves lignin degradation and increased imbibition. Germination capacity in vitro was quantified with 1 U filter-sterilised laccase added to agar medium following autoclaving, compared to a 10% bleach solution (standard bleach surface sterilisation/scarification method used in orchid seed sowing). Lignin degradation was quantified using an optical method (phloroglucinol-HCl staining) combined with image analysis, following experimental pre-treatments involving immersion in laccase solution, distilled water (negative control) or bleach (positive control). Water uptake after experimental treatments was quantified as the proportion of seeds exhibiting visible uptake of an aqueous fluorochrome under UV excitation. Laccase stimulated a doubling of germination in vitro with respect to bleach surface sterilisation/scarification alone, from 23.7 to 49.8% (P = 0.007). Laccase and bleach methods both significantly decreased the optical signal of phloroglucinol (for laccase, to 79.9 ± 1.3% of controls; anova: F = 10.333, P = 0.002). Laccase resulted in a modest but highly significant (P < 0.0001) increase in water uptake with respect to the control (11.7%; cf 99.4% for bleach). Laccase scarification can stimulate germination of A. morio through a mechanism of targeted seed coat degradation. The results demonstrate the potential of this relatively non-invasive enzymatic scarification technique

Circadian Rhythms in Visual Responsiveness in the Behaviorally Arrhythmic Drosophila Clock Mutant ClkJrk

An organism's biological day is characterized by a pattern of anticipatory physiological and behavioral changes that are governed by circadian clocks to align with the 24-h cycling environment. Here, we used flash electroretinograms (ERGs) and steady-state visually evoked potentials (SSVEPs) to examine how visual responsiveness in wild-type Drosophila melanogaster and the circadian clock mutant ClkJrk varies over circadian time. We show that the ERG parameters of wild-type flies vary over the circadian day, with a higher luminance response during the subjective night. The SSVEP response that assesses contrast sensitivity also showed a time-of-day dependence, including 2 prominent peaks within a 24-h period and a maximal response at the end of the subjective day, indicating a tradeoff between luminance and contrast sensitivity. Moreover, the behaviorally arrhythmic ClkJrk mutants maintained a circadian profile in both luminance and contrast sensitivity, but unlike the wild-types, which show bimodal profiles in their visual response, ClkJrk flies show a weakening of the bimodal character, with visual responsiveness tending to peak once a day. We conclude that the ClkJrk mutation mainly affects 1 of 2 functionally coupled oscillators and that the visual system is partially separated from the locomotor circadian circuits that drive bouts of morning and evening activity. As light exposure is a major mechanism for entrainment, our work suggests that a detailed temporal analysis of electrophysiological responses is warranted to better identify the time window at which circadian rhythms are most receptive to light-induced phase shifting

Modeling and mapping isotopic patterns in the Northwest Atlantic derived from loggerhead sea turtles

Stable isotope analysis can be used to infer geospatial linkages of highly migratory species. Identifying foraging grounds of marine organisms from their isotopic signatures is becoming de rigueur as it has been with terrestrial organisms. Sea turtles are being increasingly studied using a combination of satellite telemetry and stable isotope analysis; these studies along with those from other charismatic, highly vagile, and widely distributed species (e.g., tuna, billfish, sharks, dolphins, whales) have the potential to yield large datasets to develop methodologies to decipher migratory pathways in the marine realm. We collected tissue samples (epidermis and red blood cells) for carbon (delta C-13) and nitrogen (delta N-15) stable isotope analysis from 214 individual loggerheads (Caretta caretta) in the Northwest Atlantic Ocean (NWA). We used discriminant function analysis (DFA) to examine how well delta C-13 and delta N-15 classify loggerhead foraging areas. The DFA model was derived from isotopic signatures of 58 loggerheads equipped with satellite tags to identify foraging locations. We assessed model accuracy with the remaining 156 untracked loggerheads that were captured at their foraging locations. The DFA model correctly identified the foraging ground of 93.0% of individuals with a probability greater than 66.7%. The results of the external validation (1) confirm that assignment models based on tracked loggerheads in the NWA are robust and (2) provide the first independent evidence supporting the use of these models for migratory marine organisms. Additionally, we used these data to generate loggerhead-specific delta C-13 and delta N-15 isoscapes, the first for a predator in the Atlantic Ocean. We found a latitudinal trend of delta C-13 values with higher values in the southern region (20-25 degrees N) and a more complex pattern with delta N-15, with intermediate latitudes (30-35 degrees N) near large coastal estuaries having higher delta N-15-enrichment. These results indicate that this method with further refinement may provide a viable, more spatially-explicit option for identifying loggerhead foraging grounds

MFGE8 does not influence chorio-retinal homeostasis or choroidal neovascularization in vivo

Purpose: Milk fat globule-epidermal growth factor-factor VIII (MFGE8) is necessary for diurnal outer segment phagocytosis and promotes VEGF-dependent neovascularization. The prevalence of two single nucleotide polymorphisms (SNP) in MFGE8 was studied in two exsudative or “wet” Age-related Macular Degeneration (AMD) groups and two corresponding control groups. We studied the effect of MFGE8 deficiency on retinal homeostasis with age and on choroidal neovascularization (CNV) in mice. Methods: The distribution of the SNP (rs4945 and rs1878326) of MFGE8 was analyzed in two groups of patients with “wet” AMD and their age-matched controls from Germany and France. MFGE8-expressing cells were identified in Mfge8+/− mice expressing ß-galactosidase. Aged Mfge8+/− and Mfge8−/− mice were studied by funduscopy, histology, electron microscopy, scanning electron microscopy of vascular corrosion casts of the choroid, and after laser-induced CNV. Results: rs1878326 was associated with AMD in the French and German group. The Mfge8 promoter is highly active in photoreceptors but not in retinal pigment epithelium cells. Mfge8−/− mice did not differ from controls in terms of fundus appearance, photoreceptor cell layers, choroidal architecture or laser-induced CNV. In contrast, the Bruch's membrane (BM) was slightly but significantly thicker in Mfge8−/− mice as compared to controls. Conclusions: Despite a reproducible minor increase of rs1878326 in AMD patients and a very modest increase in BM in Mfge8−/− mice, our data suggests that MFGE8 dysfunction does not play a critical role in the pathogenesis of AMD