Patterns of phylogenetic beta diversity in China's grasslands in relation to geographical and environmental distance

Patterns and determinants of beta (β-) diversity can be used to explore the underlying mechanisms regulating community assembly. Despite being the most commonly used measure of β-diversity, species turnover does not consider the evolutionary differences among species, treating all species equally. Incorporating information on phylogenetic non-independence or relatedness among species in the calculation of β-diversity may substantially advance our understanding of the ecological and evolutionary mechanisms structuring communities. Here, we investigate the relative influence of geographical distance and differences in environmental conditions (environmental distance) on the phylogenetic β-diversity between grassland communities expanding 4000 km across the Tibetan Plateau, the Inner Mongolia Plateau and the Xinjiang Autonomous Region in China. Both observed and standardized effect size of phylogenetic β-diversity were significantly correlated with geographical and environmental distance across all regions. However, the effect of geographical distance on the standardized effect size of phylogenetic β-diversity disappeared when environmental distance was controlled. We also found that within different regions, the effect of environmental distance on both observed and standardized effect size of phylogenetic β-diversity was more significant than geographical distance. Among environmental variables, climate played a more important role in shaping observed phylogenetic β-diversity across and within regions, and standardized effect size of phylogenetic β-diversity across regions. Soil properties played a more important role in shaping standardized effect size of phylogenetic β-diversity within regions. The phylogenetic β-diversity of species from dicot and monocot clades exhibited similar patterns along environmental and geographical distance. The results suggest that at the study scale, phylogeny of grassland communities in China is predominantly structured by environmental filtering, and the dominant environmental factors may be scale-dependent.     

Taxonomic and functional diversity of stream invertebrates along an environmental stress gradient

Anthropogenic stress has been identified as main driver of freshwater biodiversity loss. Adverse effects on the biodiversity of freshwater organisms, such as macroinvertebrates, may propagate to associated ecosystem functions, such as organic matter breakdown (OMB). In this context, the functional diversity (FD) of communities has been suggested to be a more suitable predictor of changes in ecosystem functions than taxonomic diversity (TD). We investigated the response of TD and FD of invertebrate communities to an environmental stress gradient and the relation of both metrics to the rate of organic matter breakdown. For this, we sampled macroinvertebrates and determined OMB using leaf bags along an environmental stress gradient (i.e. changes in physicochemical and hydromorphological conditions) in 29 low-order streams. Taxonomic richness decreased with increasing environmental stress (r = −0.55) but was not related to OMB. Conversely, the Simpson diversity of communities was not associated with the gradient but correlated moderately (r = 0.41) with OMB. Of three functional diversity indices (functional richness, evenness and divergence), only functional richness correlated moderately with the stress gradient (r = −0.41) and any of the indices correlated with OMB. Nevertheless, functional metrics such as specific trait modalities and the total abundance of the dominant shredders correlated higher (r = 0.46 and 0.48) with OMB than the TD indices. Given a relatively small species pool in our study and methodical constraints such as the limited resolution of autecological information, the FD might perform better in other contexts and if focusing on response and effect traits for the stressor and ecosystem function under scrutiny, respectively.     

Comparing the sensitivity of diverse macroinvertebrate metrics to a multiple stressor gradient in Mediterranean streams and its influence on the assessment of ecological status

Biological indicators are increasingly being used as integrative measures of the ecosystem health in streams, particularly those using macroinvertebrate assemblage composition. Monitoring biological quality of rivers has not a long tradition in some Mediterranean European countries like Spain. Several macroinvertebrate metrics have been recently proposed to assess ecological status in Mediterranean streams, so it is necessary to compare the use of proposed biological quality metrics to select the most appropriate ones.In the present work, two classic richness metrics (total number of families and number of the Ephemeroptera, Plecoptera and Trichoptera families), three indices (IBMWP, IASPT and t-BMWQ) and two multimetric indices, recently proposed to be used in Mediterranean streams (ICM-9 and ICM-11a or IMMi-L), were compared by the analysis of the sensitivity of these metrics to a multiple stressor gradient which reflected the main pressures present in the study area. For this purpose, data from 193 sites sampled in spring (95 reference sites and 98 disturbed sites) belonging to five different Mediterranean stream types present in 35 basins were studied.The results showed that the adjusted regression coefficients (r²) for all seven metrics in the exponential regression models were higher than linear ones, thus indicating an exponential relationship between metrics and the environmental alteration. The two studied multimetric indices presented higher regression coefficients (r² = 0.590–0.669) than the three indices (r² = 0.524–0.574) and the two metrics (r² = 0.471–0.525), therefore showing a better response to a stressor gradient in Mediterranean streams. Within the multimetric indices group, ICM-11a showed the highest regression coefficients. Based on the results obtained, we suggest using the ICM-11a, apart from the IBMWP, to assess ecological status in Mediterranean streams.     

The tempo-spatial variations of phytoplankton diversities and their correlation with trophic state levels in a large eutrophic Chinese lake

Lake Chaohu is one of the most eutrophic lakes in China. Research on this lake's seasonal and spatial variations in phytoplankton diversity is needed to understand the distribution of eutrophication, as well as to find appropriate comprehensive biodiversity indices to assess the eutrophication status of the lake. The present study indicated that the Margalef index of all samples was as low as 0.799 ± 0.543 in summer (August 2011) and as high as 1.467 ± 0.653 in winter (February 2012). The Margalef index of the river samples had a high mean value and substantial variation compared with the lake samples. The Peilou index of the lake samples was higher than that of the river samples in summer and autumn (November 2011) but lower than that of the river samples in winter. In spring (May 2012), the Peilou index of the western samples was lower than that of the eastern samples. The spatial distribution of the Shannon–Wiener index was more similar to that of the Peilou index in autumn and winter, while in spring and summer, the spatial distribution was affected by both species richness and evenness. High eutrophication levels occurred in the western lake in spring and summer, whereas high levels occurred in the eastern lake, especially in the middle of the lake, in autumn and winter. The total trophic state index (TSI) in all samples exhibited a significant negative correlation with the Margalef (r = −0.726) and Peilou (r = −0.530) indices but a significant positive correlation with the Shannon–Wiener (r = 0.654) index. The partial correlation analysis results implied that these phytoplankton biodiversity indices could serve as synthetic ecological indicators to assess the eutrophication condition of Lake Chaohu.     

Ecological indicators and functional groups of copepod assemblages

Copepods are important ecological indicators of ecosystem functioning. In this study, we analyzed the structure of copepod assemblages and cross-shelf patterns based on functional traits and indicator value analysis. Copepod samples were collected from 54 stations distributed along the inner, middle, and outer continental shelves covering a wide geographical area (20,100 km²). Overall, 38 species were identified: 20 Calanoida, 14 Cyclopoida, and 4 Harpacticoida. Copepod density was the highest in the inner shelf profile, with richness and diversity being higher on the outer shelf close to the shelf break. The results suggest that there is a gradient of zooplankton distribution even on narrow (<50 km) continental shelves. The cluster analysis of the stations showed a tendency to form three groups largely corresponding to physical location. Two major functional groups were identified, sorted by spawning strategy. These were subdivided according to the trophic regime and feeding strategy. Temora turbinata (alien species), Clausocalanus furcatus, and Ditrichocorycaeus amazonicus (native species) were found to be ecological indicators. These species have different functional traits and distinct cross-shelf distributions, where the alien species dominates the turbid coastal waters, and C. furcatus is an indicator of outer-shelf waters. Thus, these species can be considered ecological indicators of the different continental shelf waters. The results indicate that it is necessary to understand the functional diversity and ecological indicators of copepods, considering their importance for the basis of marine food webs.     

Pod,seed traits and cytotaxonomic studies of some Vicia narbonensis L. accessions (Fabaceae)

This study aimed to characterize eight accessions of Vicia narbonensis L. originated from different Mediterranean countries. The cytology of these species is rarely known despite the fact of its great socio-economical and ecological interest in these arid and semi-arid zones. This work aimed mainly to characterize the karyotype, morphological pod and seed traits of the species. Karyotypes of all accessions were similar to a diploid number of 2n = 2x = 14. All the accessions have submetacentric chromosomes with a secondary constriction attached to the long arm of pair VII close to the centromere. Variation in chromosome size was observed; it ranged from 5.86 μm to 7.62 μm. Indices of karyotype asymmetry were calculated as the total form percentage (TF%) and symmetric indices (Syi) which ranged from 33.75% to 35.42% and from 51.01% to 54.85%, respectively. The predominance of submetacentric chromosomes indicated that the karyotype is symmetrical and can be considered as primitive. However, the analysis of quantitative parameters measured on pods and seeds showed a significant variation between accessions. A relationship between centromeric index and the pod beak length was found. Estimation of phenotypic diversity using the Shannon diversity index (H′) showed that the length, the seed color and the number of seeds per pod are the most polymorphic traits with respectively, H′ = 0.92, 0.80 and 0.83. Cluster analysis of karyological, pod and seed traits showed four groups of accessions. This clustering is partially due to the geographical origin of the studied accessions. The variation in chromosome size, pod and seed traits could offer potentially valuable genetic resources for the improvement of V. narbonensis which is considered as neglected and underutilized crop species (NUCS).     

Spatial distribution of soil penetration resistance as affected by soil compaction: The fractal approach

Using semivariograms and fractal dimension (D), we identified the spatial variation of penetration resistance (PR) in variously compacted silty loam in the Lublin region, South-East Poland. Four compaction treatments were as follows: zero traffic (0p), one pass of tractor (1p), three passes of tractor (3p), and (8p) eight passes. Penetration resistance was measured in a square net of 0.6 m × 0.6 m with grid density of 0.05 m, in horizontal planes at depths of 0.05, 0.15, 0.25, 0.35, 0.45 and 0.55 m in each traffic treatment. The data were analysed in 6 horizontal planes and 12 vertical planes. The vertical planes were obtained by transforming the measured data along one side of the square at every 0.05 m. Total number of penetrations was 864. Fractal dimension (D) was estimated from the slope of the log–log semivariogram plots. The semivariograms showed spatial autocorrelation of penetration resistance in the horizontal and vertical planes. Direction in space was important in this study. In the horizontal planes the differentiation of penetration resistance semivariance at different depths was considerable and not clearly related with traffic intensity. In the vertical planes the semivariograms showed spatial dependence of the PR and evident decreasing of semivariance with increasing traffic intensity. Kriging-interpolated maps revealed that the differentiation of penetration resistance was higher in the vertical than in the horizontal planes. The vertical differentiation was higher in 0p than in the remaining treatments. The overall mean fractal dimensions in the vertical planes increased with increasing compaction levels and can be a useful indicator of the compaction level. The opposite courses of fractal dimension in the vertical and horizontal planes indicate spatial anisotropy in distribution of penetration resistance.     

A multivariate analysis integrating ecological,socioeconomic and physical characteristics to investigate urban forest cover and plant diversity in Beijing,China

Understanding the factors driving the variation in urban green space and plant communities in heterogeneous urban landscapes is crucial for maintaining biodiversity and important ecosystem services. In this study, we used a combination of field surveys, remote sensing, census data and spatial analysis to investigate the interrelationships among geographical and social-economic variables across 328 different urban structural units (USUs) and how they may influence the distributions of urban forest cover, plant diversity and abundance, within the central urban area of Beijing, China. We found that the urban green space coverage varied substantially across different types of USUs, with higher in agricultural lands (N = 15), parks (N = 46) and lowest in utility zones (N = 36). The amount of urban green space within USUs declines exponentially with the distance to urban center. Our study suggested that geographical, social and economic factors were closely related with each other in urban ecological systems, and have important impacts on urban forest coverage and abundance. The percentage of forest as well as high and low density urban areas were mainly responsible for variations in the data across all USUs and all land use/land cover types, and thus are important constituents and ecological indicators for understanding and modeling urban environment. Herb richness is more strongly correlated with tree and shrub density than with tree and shrub richness (r = −0.472, p < 0.05). However, other geographic and socioeconomic factors showed no significant relationships with urban plant diversity or abundance.     

Effects of plant diversity on microbial biomass and community metabolic profiles in a full-scale constructed wetland

There has been less understanding of relations of microbial community patterns with plant diversity in constructed wetlands. We conducted a single full-scale subsurface vertical flow constructed wetland (SVFCW, 1000 m²) study focusing on domestic wastewater processing. This study measured the size and structure of microbial community using fumigation extraction and BIOLOG Ecoplate? techniques, to examine the effects of macrophyte diversity on microbial communities that are critical in treatment efficiency of constructed wetlands. We also determined the relationship of plant diversity (species richness) with its biomass production under disturbance of the same wastewater supply. Linear regression analysis showed that plant biomass production strongly correlated with plant species richness (R = 0.407, P < 0.001). Increase in plant species richness increased microbial biomass carbon and nitrogen (R = 0.494, P < 0.001; R = 0.465, P < 0.001) and utilization of amino acids on Ecoplates (R = 0.235, P = 0.03), but limited the utilization of amine/amides (R = ?0.338, P = 0.013). Principal components analysis (PCA) showed that the diversity and community-level physiological profiles (CLPP) of microbial community at 168 h of incubation strongly depended on the presence or absence of plant species in the SVFCW system, but not on the species richness. This is the first step toward understanding relations of plant diversity with soil microbial community patterns in constructed wetlands, but the effect of species diversity on microbial community should be further studied.     

Sea-surface temperature for left-coiling Neogloboquadrina populations inhabiting the westernmost Mediterranean in the middle Pleistocene and the Pleistocene-Pliocene transition

Using the modern analog methodology applied to planktonic foraminifers, we analyze the relation between the frequency of the coiling type in Neogloboquadrina populations and the sea-surface temperatures (SST) during the middle Pleistocene and the Pleistocene-Pliocene transition in the Alboran Sea (westernmost Mediterranean), close to the Atlantic connection. The results reveal that the present-day positive correlation between the two variables (r = 0.649) is maintained even with a higher coefficient (r = 0.783) in the middle Pleistocene but falls slightly (r = 0.517) in the Pleistocene-Pliocene transition due mainly to a dispersal of the temperatures for the samples bearing predominantly left-coiling Neogloboquadrina. The temperature used as a reference for the coiling change resulted 5-6 °C above its North Atlantic present-day reciprocal, but these thermal differences could be caused by: i) a slight overestimate (1–2 °C) due to the lumping of all neogloboquadrinids into a single variable when SST are estimated; ii) only the warmer range of temperatures for the left-coiling populations would be represented in the samples; and iii) a remarkable warming inside the westernmost Mediterranean during the summer stage mixing cold and warm assemblages in bottom sediments. In addition, these results in combination with those derived from the isotopic analyses (δ¹⁸O) in G. bulloides tests, suggest that during the Pliocene-Pleistocene transition two different populations of left-coiling Neogloboquadrina could have existed with different environmental requirements: one, derived from late Miocene-Pliocene (i.e., left-coiling N. acostaensis group), and another being the ancestor of the modern N. pachyderma (left-coiling). Similarity analyses were achieved in order to locate the position of the core-tops with the assemblages most analogous to those of the fossil samples containing left-coiling Neogloboquadrina. The results reinforce the idea that the presence of these forms in the Mediterranean during the intervals studied would be related mainly to the input of cold waters from the North Atlantic during glacial stages, although it could be secondarily favored by the establishment of upwelling conditions, as in the present-day North Alboran waters.     

Genetic diversity and population structure in Vallisneria spinulosa (Hydrocharitaceae)

Vallisneria spinulosa is a dominant submerged macrophyte in lakes of the middle–lower reaches of the Yangtze River. Allozyme variation, clonal diversity and population genetic structure were investigated for a total of 396 individuals sampled from 10 extant populations. V. spinulosa maintained high levels of genetic variation both at the species (P = 46.2, A = 1.69, H_e = 0.23) and at the population level (P = 46.2, A = 1.58, H_e = 0.21). Although aquatic macrophytes commonly exhibit low genetic variation within populations, the obligately outcrossing mating system of V. spinulosa and pervasive gene flow likely account for the high levels of diversity maintained within populations. All V. spinulosa populations contained high clonal diversity with a mean proportion of distinguishable genotypes of 0.57 and a mean Simpson's diversity index of 0.95, indicating that populations were founded sexually or that successful seedling recruitment occurred after initial colonization. Partitioning of genetic diversity revealed a surprisingly low population differentiation (G_ST = 0.06) as compared to other hydrophilous angiosperms. No evidence of isolation-by-distance was found (r = 0.056, P = 0.312), suggesting that gene flow was not restricted geographically. The UPGMA cluster analysis revealed that several widely separated populations grouped together, suggesting long-distance gene flow among populations. The high vagility of V. spinulosa and extensive hydrologic connectivity among populations have facilitated long-distance gene flow and resulted in the pattern of population genetic structure in V. spinulosa.     

Relationship between zinc levels and plasma leptin in hemodialysis patients

Recent evidences suggested a possible relationship between zinc deficiency and leptin levels in pathogenesis of anorexia in chronic kidney disease. The present study addressed the relationship between zinc and leptin in hemodialysis (HD) patients.MethodsFifty HD patients (54.3 ± 12.7 years old, 62% men) were studied and compared to 21 healthy volunteers (50.7 ± 15.7 years old, 43% men). Biochemical data, serum zinc, plasma leptin, IL-6, TNF-α and C-Reactive Protein levels were determined. Anthropometric parameters, food intake and appetite score were also assessed.ResultsThe leptin levels were higher in HD patients (16.1 μg/mL (0.21–118.25) vs 6.0 μg/mL (0.50–23.10)) in healthy volunteers (p = 0.04), whereas serum zinc levels were lower (54.5 ± 16.3 μg/dL) compared to healthy volunteers (78.4 ± 9.4 μg/dL) (p = 0.0001). The plasma leptin was correlated negatively with plasma zinc (r = ?0.33; p = 0.007), energy (r = ?0.38; p = 0.002) and protein intake (r = ?0.34; p = 0.006) and, positively correlated with BMI (r = 0.54; p = 0.0001), % body fat (r = 0.70; p = 0.0001) and conicity index (r = 0.46; p = 0.001). Plasma zinc was associated with hemoglobin (r = 0.30; p = 0.04) and negatively associated with TNF-α (r = ?0.37; p = 0.002) and C-Reactive Protein (r = ?0.37; p = 0.004). There was no correlation among Zn, leptin and appetite score in these patients.ConclusionThis study showed that low plasma zinc levels are negatively associated with high leptin levels in HD patients.     

Kinetic modeling and implementation of superior process strategies for β-galactosidase production during submerged fermentation in a stirred tank bioreactor

This paper reports development and implementation of superior fermentation strategies for β-galactosidase production by Lactobacillus acidophilus in a stirred-tank bioreactor. Process parameters (aeration and agitation) were optimized for the process by application of Central Composite Design. Aeration rate of 0.5 vvm and agitation speed of 250 rpm were most suitable for β-galactosidase production (2001.2 U/L). Further improvement of the operation in pH controlled environment resulted in 2135 U/L of β-galactosidase with productivity of 142.39 U/L h. Kinetic modeling for biomass and enzyme production and substrate utilization were carried out at the aforementioned pH controlled conditions. The logistic regression model (X₀ = 0.01 g/L; X_max = 2.948 g/L; μ_max = 0.59/h; R² = 0.97) was used for mathematical interpretation of biomass production. Mercier's model proved to be better than Luedeking–Piret model in describing β-galactosidase production (P₀ = 0.7942 U/L; P_max = 2169.3 U/L; P_r = 0.696/h; R² = 0.99) whereas the latter was more efficient in mathematical illustration of lactose utilization (m = 0.187 g/g h; Y_x/s = 0.301 g/L; R² = 0.98) among the two used in this study. Strategies like fed-batch fermentation (3694.6 U/L) and semi-continuous fermentation (5551.9 U/L) further enhanced β-galactosidase production by 1.8 and 2.8 fold respectively.     

Scattered trees and livestock grazing as keystones organisms for sustainable use and conservation of Mediterranean dehesas

Mediterranean scattered oak woodlands support relatively high biological diversity, and provide important ecosystem services. However, there is still a lack of knowledge about livestock-tree-grassland relationships, knowledge necessary for effective conservation management in these systems. To address such lacuna, we measured the effects of scattered trees and grazing animals (livestock vs. wild ungulates, mostly deer) on biomass and diversity (and their relationship) of the herbaceous layer (understory) of open holm oak woodland (known as dehesa) in Central Spain, for two years. We located two 1-ha plots within three different grazing management schemes (cattle, sheep and wildlife only). Within each plot, we randomly selected four focal trees from which a subplot was established in two directions and three distances. In each subplot, we measured plant diversity and biomass, along with microclimatic variables across sites. We found rainfall variability affected herbaceous biomass and diversity in the dehesa system, and the effects were nuanced: in a dry year (<300 mm annual rainfall) the presence of livestock grazing correlated with higher herbaceous biomass (14%), and in moister year (>600 mm annual rainfall) the effect was enhanced, as plots under livestock grazing exhibited even higher herbaceous biomass (42%). In addition, livestock, particularly cattle, generated a more diverse community (52 species m⁻²). Microsites created by trees generated high plant diversity among herbaceous communities, where alpha diversity was higher (20%) in ecotone and inter-tree gaps than under the canopies. In addition, species turnover was higher than 50% in all cases. Biomass was higher (15%) under the canopies only under humid conditions. Legume dry matter was positively related to plant diversity. We concluded that agroforestry management practices such as promoting tree development through silvicultural techniques to create different microsites, and maintaining livestock grazing are needed to preserve these emblematic ecosystems.     

Segmentation d’images de lésions cérébrales primitives en TEP FET : influence du volume de travail

Purpose(1) Evaluate the reproducibility of segmentation methods depending on the preselection region for tumour volume determination on ¹⁸F-fluoro-ethyl-tyrosine (FET) PET. (2) Evaluate the intra and inter-operator reproducibility of the manual delineation. (3) Compare this delineation with the segmentation methods.Materials and methodsEighteen FET PET of patients with glioblastoma were analysed. Preselection regions were determined prior to any segmentation. Two physicians delineated the tumour volume manually. The tumour volume was also delineated with a threshold method (40 and 70% of SUV_max), and a random walk based method. Pearson coefficient (r) (P < 0.05 for r > 0.468) and Jaccard indices (JI) were used to compare the volumes.ResultsManual delineation was reproducible with r = 0.97 and IJ = 0.65 for intra-operator, and r = 0.76 and IJ = 0.45 for inter-operator reproducibility. The preselection regions for a given lesion were different and the segmentation varied with the preselection region: r = 0.55 JI = 0.58; r = 0.85 JI = 0.83; r = 0.70 JI = 0.39 respectively for the threshold of 40%, 70% and the random walk. The segmentation differed form de manual delineation with r = 0.37 and JI = 0.16; r = 0.54 and JI = 0.42; r = 0.43 and JI = 0.37 respectively for the threshold of 40%, 70% and the random walk.ConclusionThe reproducibility of the segmentation methods depends extensively on the preselection region. The intra-operator reproducibility of cerebral lesion delineation on FET PET is satisfactory. The inter-operator reproducibility could be improved.     

Immunoglobulin genes in Andalusia (Spain). Genetic diversity in the Mediterranean space

Andalusia is the most densely populated region of Spain since ancient times, and has a rich history of contacts across the Mediterranean. Earlier studies have underlined the relatively high frequency of the Sub-Saharan GM 1,17 5* haplotype in western Andalusia (Huelva province, n = 252) and neighbouring Atlantic regions. Here, we provide novel data on GM/KM markers in eastern Andalusians (n = 195) from Granada province, where African GM*1,17 5* frequency is relatively high (0.044). The most frequent GM haplotypes in Andalusia parallel the most common in Europe. Altogether, these data allow us to gain insight into the genetic diversity of southern Iberia. Additionally, we assess population structure by comparing our Iberian samples with 41 Mediterranean populations. GM haplotype variation across the Mediterranean reflects intense and complex interactions between North Africans and South Europeans along human history, highlighting that African influence over the Iberian Peninsula does not follow an isotropic pattern.     

Estimating Pliocene sea-surface temperatures in the Mediterranean: An approach based on the modern analogs technique

The possibilities of adapting the Modern Analogs Technique (MAT) based on planktonic foraminifers for estimating sea-surface temperatures (SST) in the Mediterranean during the Pliocene are discussed in this article. The calibration database used comprises 684 core-top samples distributed in the North Atlantic and the Mediterranean. MAT estimates show an imperceptible bias (− 0.03 °C; σ = 0.59) and a low mean error of estimates (0.42 °C; σ = 0.42) when applied over the samples of the calibration dataset. The procedure used for assimilating the Pliocene taxonomic categories to those of the modern assemblages results in an increase from 17 to 40 in the number of samples showing an error > 2 °C when applied over the calibration database. However, the precision of MAT does not diminish when these samples are removed from the dataset. This methodology was used for obtaining SST estimates of late-middle Pliocene–earliest Pleistocene samples from ODP-site 975 (Menorca area), which have close modern analogs within the calibration database. In order to compare this technique with an additional proxy, we measured also δ¹⁸O values of G. bulloides tests from these samples. The results obtained show a good agreement on the whole, which corroborates the validity of the technical approach proposed.