ALBERT

Description: Methane is a major contributor to the greenhouse effect, its atmospheric concentration being more than doubled since the XIX century. Every year 22 Tg of methane are released to the atmosphere from several natural and anthropogenic sources. Natural sources include geothermal/volcanic areas but the estimation of the total methane emission from these areas is currently not well defined since the balance between emission through degassing and microbial oxidation within the soils is not well known. Microbial oxidation in soils contributes globally for about 3-9% to the removal of methane from the atmosphere and recent studies evidenced methanotrophic activity also in soils of volcanic/geothermal areas despite their harsh environmental conditions (high temperatures, low pH and high concentrations of H2S and NH3). Methanotrophs are a diverse group of bacteria that are able to metabolize methane as their only source of carbon and energy and are found within the Alpha and Gamma classes of Proteobacteria and within the phylum Verrucomicrobia. Our purpose was to study the interaction between methanotrophic communities and the methane emitted from the geothermally most active site of Pantelleria island (Italy), Favara Grande, whose total methane emission has been previously estimated in about 2.5 t/a. Laboratory incubation experiments with soil samples from Favara Grande showed methane consumption values of up to 9500 ng g-1 dry soil per hour while soils collected outside the geothermal area consume less than 6 ng g-1 h-1. The maximum consumption was measured in the shallowest part of the soil profile (1-3 cm) and high values (〉100 ng g-1 h-1) were maintained up to a depht of 15 cm. Furthermore, the highest consumption was measured at 37 C, and a still recognizable consumption (〉20 ng g-1 h-1) at 80 C, with positive correlation with the methane concentration in the incubation atmosphere. These results can be considered a clear evidence of the presence of methanotrophs that were investigated by culturing and culture-independent techniques. The diversity of proteobacterial methanotrophs was investigated by creating a clone library of the amplified methane mono-oxygenase encoding gene, pmoA. Clone sequencing indicates the presence of Gammaproteobacteria in the soils of Favara Grande. Enrichment cultures, on a mineral medium in a CH4-enriched atmosphere, led to the isolation of different strains that were identified as Methylocistis spp., which belong to the Alphaproteobacteria. The presence of Verrucomicrobia was detected by amplification of pmoA gene using newly designed primers. Soils from Favara Grande show therefore the largest spectrum of methanotrophic microorganisms until now detected in a geothermal environment. While the presence of Verrucomicrobia in geothermal soils was predictable due to their thermophilic and acidophilic character, the presence of both Alpha and Gamma proteobacteria was unexpected. Their presence is limited to the shallowest part of the soil were temperatures are lower and is probably favored by a soil pH that is not too low (pH 5) and their contribution to biological methane oxidation at Pantelleria is significant. Understanding the ecology of methanotrophy in geothermal sites will increase our knowledge of the role of soils in methane emissions in such environments.

Description: Published

Description: Vienna, Austria

Description: 4.5. Studi sul degassamento naturale e sui gas petroliferi

Description: open

Description: Methane plays an important role in the Earth’s atmospheric chemistry and radiative balance being the most important greenhouse gas after carbon dioxide. It has recently been established that geogenic gases contribute significantly to the natural CH4 flux to the atmosphere (Etiope et al., 2008). Volcanic/geothermal areas contribute to this flux, being the site of widespread diffuse degassing of endogenous gases (Chiodini et al., 2005). In such an environment soils are a source rather than a sink for atmospheric CH4 (Cardellini et al., 2003; Castaldi and Tedesco, 2005; D’Alessandro et al., 2009; 2011; 2013). Due to the fact that methane soil flux measurements are laboratory intensive, very few data have been collected until now in these areas. Preliminary studies (Etiope et al., 2007) estimated a total CH4 emission from European geothermal and volcanic systems in the range 4-16 kt a-1. This estimate was obtained indirectly from CO2 or H2O output data and from CO2/CH4 or H2O/CH4 values measured in the main gaseous manifestations. Such methods, although acceptable to obtain order-of-magnitude estimates, completely disregard possible methanotrophic activity within the soil. At the global scale, microbial oxidation in soils contributes for about 3-9% to the total removal of methane from the atmosphere. But the importance of methanotrophic organisms is even larger because they oxidise the greatest part of the methane produced in the soil and in the subsoil before its emission to the atmosphere. Environmental conditions in the soils of volcanic/geothermal areas (i.e. low oxygen content, high temperature and proton activity, etc.) have been considered inadequate for methanotrophic microrganisms. But recently, it has been demonstrated that methanotrophic consumption in soils occurs also under such harsh conditions due to the presence of acidophilic and thermophilic Verrucomicrobia. These organisms were found in Italy at the Solfatara di Pozzuoli (Pol et al., 2007), in New Zealand at Hell’s Gate (Dunfield et al., 2007) and in Kamchatka, Russia (Islam et al., 2008). Both the Italian and the Hellenic territories are geodynamically very active with many active volcanic and geothermal areas. Here we report on methane flux measurements made at Pantelleria (Italy) and at Sousaki and Nisyros (Greece). The total methane output of these three systems is about 10, 19 and 1 t a-1, respectively (D’Alessandro et al., 2009; 2011; 2013). The total emissions obtained from methane flux measurements are up to one order of magnitude lower than those obtained through indirect estimations. Clues of methanotrophic activity within the soils of these areas can be found in the CH4/CO2 ratio of the flux measurements which is always lower than that of the respective fumarolic manifestations, indicating a loss of CH4 during the travel of the gases towards earth’s surface. Furthermore laboratory methane consumption experiments made on soils collected at Pantelleria and Sousaki revealed, for most samples, CH4 consumption rates up to 9.50 µg h-1 and 0.52 µg h-1 respectively for each gram of soil (dry weight). Only few soil samples displayed no methane consumption activity. Finally, microbiological and molecular investigations allowed us to identify the presence of methanotrophic bacteria belonging to the Verrucomicrobia and to the Alpha- and Gamma-Proteobacteria in the soils of the geothermal area of Favara Grande at Pantelleria. While the presence of the former was not unexpected due to the fact that they include acidophilic and thermophilic organisms that were previously found in other geothermal environments, the latter are generally considered not adapted to live in harsh geothermal environments. Their presence in the soils of Pantelleria could be explained by the fact that these soils do not have extremely low pH values (〉5). Indeed thermotollerant methanotrophic Gamma-proteobacteria, have been previously found in the sediments of thermal springs in Kamchatka (Kizilova et al., 2012). Such species could find their niches in the shallowest part of the soils of Favara Grande were the temperatures are not so high and they thrive on the abundant upraising hydrothermal methane.

Description: Published

Description: Patras, Greece

Description: 4.5. Studi sul degassamento naturale e sui gas petroliferi

Description: open

Description: Methane plays an important role in the Earth’s atmospheric chemistry and radiative balance being the most important greenhouse gas after carbon dioxide. It is released to the atmosphere by a wide number of sources, both natural and anthropogenic, with the latter being twice as large as the former. It has recently been established that significant amounts of geological methane, produced within the Earth’s crust, are currently released naturally into the atmosphere. Active or recent volcanic-geothermal areas represent one of these sources of geological methane. Due to the fact that methane flux measurements are laboratory intensive, very few data have been collected until now and the contribution of this source has been often indirectly estimated. Both the Italian and the Hellenic territories are geodynamically very active with many volcanic and geothermal areas. Here we report on methane flux measurements made at Pantelleria (Italy), Sousaki and Nisyros (Greece). The total outputs of these three systems are about 10, 19 and 2 t a-1 respectively. These figures are up to one order of magnitude lower than those obtained through indirect estimations. At the global scale, microbial oxidation in soils contributes to the total removal of methane from the atmosphere. Environmental conditions in the soils of volcanic/geothermal areas (i.e. low pH, high temperature, etc.) have been considered inadequate for methanotrophic microrganisms. But recently, it has been demonstrated that methanotrophic consumption in soils occurs also under such harsh conditions due to the presence of thermo-acidophilic Verrucomicrobia. Here we present the results of laboratory incubation experiments on soil samples collected at the main exhalative areas that highlighted methanotrophic activity also at Pantelleria and Sousaki. Soil metagenomic DNA was extracted from some of the Pantelleria samples and analysed using Temporal Temperature Gradient Electrophoresis (TTGE) of the amplified Bacterial 16S rRNA gene in order to evaluate the total bacterial diversity. Soil DNA amplification with primers targeting Proteobacterial and Verrucomicrobial methane monooxygenase genes (pmmo) revealed the presence of methanotrophs affiliated to both phyla up to a depth of 11 cm and a temperature of 80°C. The diversity of proteobacterial methanotrophs was investigated by creating a clone library of the amplified methane mono-oxygenase encoding gene, pmmoA. The clone sequences are close to those of uncultured type I methanotrophic proteobacteria. An attempt to isolate methanotrophs was carried out on soils from Pantelleria, sampled at different depths, by enrichment cultures on a mineral medium in a methane-enriched atmosphere. No isolates were obtained from enrichments carried out at 65°C while incubation at 37°C allowed to isolate a few methanothropic strains that were identified as Methylocystis spp.

Description: Published

Description: Kagoshima, Japan

Description: 4.5. Studi sul degassamento naturale e sui gas petroliferi

Description: open

Description: Pantelleria is an active volcanic complex, at present in quiescent status, hosting a high enthalpy geothermal system. Explorative geothermal wells tapped exploitable water-dominated reservoirs at 600-800 m depth with maximum measured temperatures of 250°C. Five field campaigns for soil gas measurements were made in the period from July 2005 to October 2006. CO2 flux was measured with the accumulation chamber method at 807 sites, CO2 concentration and Rn activity in soil atmosphere were measured at 50 cm depth at 728 and 358 sites, respectively. The first campaign covered the whole island (about 83 km2) with an approximately sample density of 3.5 points per km2 for CO2 (flux and concentration) measurements and of 1.6 points per km2 for Rn measurements. The distribution of the sampling points was as far as possible evenly distributed. Only few limited areas resulted uncovered due to accessibility problems (inhabited areas, airport, steep or rough topography). In the following four surveys nine areas were studied with greater detail with sample densities of up to 100 points per km2. Flux measurements for the whole surveys gave values in the range 〈 0.1 - 4700 gCO2 m-2 day-1 (Fig. 1). Organic contribution could not be distinguished with statistical methods and in the present study was considered negligible above 30 g m-2 day-1. The sites displaying flux values above this threshold, representing about 32% of the total population, are almost all located within the 50 ka old caldera. The highest values correspond to the areas of Favara Grande and of the lake Specchio di Venere (Fig. 1), which have long been recognized as sites of anomalous degassing with the presence of active fumarolic vents with temperatures of about 100 °C in the former and thermal springs with abundant bubbling gases and a mofette in the latter. A new anomalous degassing area with very high output values has been identified on the southern flank of Mt. Grande (MGS in Fig. 1). In this area the vegetation cover strongly contrasts with that of the neighboring areas, consisting of scrubby low growing plants, mosses and bare land. Indeed, high CO2 flux values are always measured as long as such vegetation cover is found, abruptly decreasing when higher growing plants are present. The contrast is particularly evident in springtime and can be used as a good marker to individuate anomalous degassing areas. Further areas were studied in detail being sites of fumarolic manifestations. All of them are also sites of anomalous CO2 fluxes although of limited areal extension. The area of the last subaerial volcanic activity (Mursia) was also studied but the measurements were all far below the anomaly threshold. The total CO2 output of the anomalous degassing areas was estimated through geostatistical methods considering only values above the anomaly threshold. Results evidence that most of the output (87%) of the island is due to 3 of the anomalous degassing areas (Lake, MGS and Favare), the first of which accounts for more than 50%. Summing up the contribution of all anomalous areas we obtained a total output of about 0.3 kg s-1 (26 t day-1) over an area of about 0.58 km2. Concentrations of CO2 ranged from 0.039 (atmospheric value) up to 95 % (Fig. 1). The distribution of the values on a probability plot evidenced two statistically distinct populations with an inflection point at about 0.8 %. The values below the threshold (65 % of the population) can be considered as derived from organic activity while those above of magmatic/geothermal origin. The spatial distribution of the sites with anomalous concentrations closely resembles those of anomalous CO2 fluxes. Radon222 activity in the soil ranged from 〈 0.1 to 〉1000 kBq m-3 (Fig. 1). The distribution of the values on a probability plot evidenced three statistically distinct populations with inflection points at 40 and 400 kBq m-3. The lower population (74% of the entire population) probably corresponds to close to equilibrium values in soils with different contents of parent isotopes of the 238U decay chain and could be considered as background population. The values of the other two populations have to be considered anomalous and their high activity values have to be related either to sustained fluxes of a carrier gas (CO2) or/and to enhanced release from the soil due to fumarolic alteration. These anomalous values generally correspond to elevated CO2 flux values and are found close to active or fossil fumarolic areas. The close relationship with fumarolic alteration is confirmed by the contrasting behavior of two of the areas displaying the highest CO2 flux values, namely Favare and Lake. In the former area the high soil temperatures, testifying for anomalous fluxes of hydrothermal fluids, are reflected in a high percentage of anomalous values of soil 222Rn activity. On the contrary at the lake area, where the highest CO2 fluxes are measured, soil temperatures are much lower and consequently also soil 222Rn activity. The only high soil 222Rn activity values are measured along the southern shores of the lake where seeps of thermal water with about 60 °C are present. At Mursia, where recent basaltic lavas and scorias crop out, soil 222Rn activity is particularly low due to the low content of parent radioactive elements in these rocks. Finally, the present study evidenced a few areas were the gas hazard due to both CO2 and radon is elevated with either acute or chronic health issues for humans. These are the western shores of the lake and the village of Rekale the only inhabited area close to an anomalous degassing area.

Description: Published

Description: Patras, Greece

Description: 4.5. Studi sul degassamento naturale e sui gas petroliferi

Description: open