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    Caller ID for Risso’s and Pacific White-sided dolphins

    The Bayesian VMD Method we developed can classify pulsed signals with similar frequency content in poor SNR files from underwater acoustic recordings. The Method consists of two parts. The first part scans the incoming audio data as segments that potentially contain signals of interest by detecting energy peaks. It then uses the start and end of the energy peaks to isolate those areas of interest from non-signal areas of the audio file. The second part classifies the detected signals into separate categories based on their frequency content. The algorithms of our Detector and Classifier steps are self-developed, but some key components in them were inspired by previous work39,40,41.DetectorThe proposed detector uses full audio files that are 4.5 s long at a sampling rate of 100 kHz. It then finds audio file segments where potential signals of interest exist.For a given audio file, denoted by ({hat{x}}(n)), where (n=1, dots , N), and N is the total number of samples, the Laplacian Differential Operator (LDO) is applied to ({hat{x}}(n)) resulting in an enhanced version of the audio file denoted by y(n), as follows:$$begin{aligned} y(n) = frac{1}{4}frac{partial ^2 {hat{x}}}{partial n^2} end{aligned}$$
    (1)
    The LDO enhances the transient signals (edge detection) and filters out the low frequencies ((< 10) kHz) which are not needed for Gg and Lo pulsed signal classification. The y(n) is then transformed into a time-frequency representation using Short-time Fourier transform (STFT). The STFT was implemented on 1024 samples with 90% overlap and a 1024-point Hanning window. The magnitude of the STFT matrix s(n, f) is given as ({hat{S}}(n,f)).$$begin{aligned} {hat{S}}(n,f) = begin{bmatrix} |s_{11}| &{} dots &{} |s_{1N}|\ vdots &{} ddots \ |s_{M1}| &{} &{} |s_{MN}| end{bmatrix} end{aligned}$$ (2) Where (N) is the length of the input segment and (M) is the number of frequency bins. The dimensionality of matrix ({hat{S}}(n,f)) is reduced from 2-D to 1-D as follows:$$begin{aligned} S_{d}(n) = sum _{f=1}^{M} {hat{S}}(n,f) end{aligned}$$ (3) The resulting temporal sequence is an accumulated sum of all frequency bins from (begin{aligned} {hat{S}}(n,f) end{aligned}), so scaling is applied, as follows:$$begin{aligned} S_{d}(n) = frac{S_{d}(n)}{max{S_{d}(n)}} end{aligned}$$ (4) After finding (S_{d}(n)) from Eq. (4), the mean of (S_{d}(n)) is subtracted. Then, to determine the boundaries of the acoustic signal, an adaptive threshold is applied. The first step in developing the threshold is to vectorize the matrix ({hat{S}}(n,f)) in column order into a vector called (S_{r}(n)):$$begin{aligned} S_{r}(n) = overrightarrow{{hat{S}}(n,f)} end{aligned}$$ (5) Then, (S_r (n)) is scaled similar to (S_{d}(n)) and is sorted into ascending order, denoted by ({hat{S}}_r(n)). The changing point where the root-mean-square level of the sorted curve ({hat{S}}_r(n)) changes the most is obtained by minimizing Eq. (6)39,40,42$$begin{aligned} J(k) = sum _{i=1}^{k-1} Delta ({hat{S}}_{r,i}; chi ([{hat{S}}_{r,1} dots {hat{S}}_{r,k-1}])) + sum _{i=k}^{N} Delta ({hat{S}}_{r,i}; chi ([{hat{S}}_{r,k} dots {hat{S}}_{r,N}])) end{aligned}$$ (6) where (k) and N are the index of the changing point and the length of the sorted curve ({hat{S}}_r (n)), respectively, and$$begin{aligned} sum _{i=u}^{v} Delta ({hat{S}}_{r,i}; chi ([{hat{S}}_{r,u} dots {hat{S}}_{r,v}])) = (u-v+1)log left( frac{1}{u-v+1}sum _{n=u}^{v}{hat{S}}_{r,n},^{2}right) end{aligned}$$ (7) The threshold, (lambda), is the value of ({hat{S}}_r (k)) which equals the noise floor estimation, and can be represented as follows:$$begin{aligned} begin{aligned} {mathcal {H}}_{0}: S_d(n) < lambda \ {mathcal {H}}_{1}: S_d(n) ge lambda end{aligned} end{aligned}$$ (8) where ({mathcal {H}}_0) and ({mathcal {H}}_1) are the hypothesis that the activity was below or above the threshold, respectively. The calculated threshold can vary for each file, thus making it adaptable if ambient noise conditions change between files. The threshold (lambda) is then projected onto the temporal sequence (S_{d}(n)) to extract the boundaries of the regions of the acoustic signal that comprised the detected energy peak. The start and end points of each acoustic signal are determined as the first and last points that are greater than (lambda) in amplitude.The boundaries of the detected segments are scaled by the sampling rate to obtain start and end times which will be used to extract the audio file segments from the original data file in the classification step. Figure 4 illustrates the layout of the the proposed detector.Figure 4Block diagram of the proposed detector.Full size imageClassifierOnce segments with energy peaks were identified, they were scanned by the team’s bioacoustics expert, and any segments confirmed to contain only Gg or Lo signals were sifted out for use in testing the accuracy of the Bayesian VMD Method classifier.In this paper, the metric weight was defined for classification purposes. The weight for a parameter (varvec{theta _i}) given its measurement (varvec{y_i}) is defined as$$begin{aligned} w(varvec{theta _i} mid varvec{y_i}) = P_{varvec{Theta mid Y}}(varvec{theta _i} mid varvec{y_i}) * varvec{p_i} end{aligned}$$ (9) where (varvec{theta _i}) is the probability density function (PDF) of (varvec{y_i}), (varvec{y_i}) is one measurement in the measurement vector (varvec{y}), (P_{varvec{Theta mid Y}}(varvec{theta _i} mid varvec{y_i})) is the posterior probability of the parameter (varvec{theta _i}) given the measurement (varvec{y_i}), and (varvec{p_i}) is the scaled prominence value of (varvec{y_i}).When a detected audio file segment is fed into the Bayesian VMD classifier, the classification process starts with a feature extraction step. During this step, peak and notch frequencies and their prominence values were obtained from the VMD-Hilbert spectrum of the segment. The prominence measures how much a peak stands out due to its intrinsic height or how much a notch stands out due to its depth and its location relative to surrounding peaks or notches. In general, peaks that are taller and more isolated have a higher “prominence” (p) than peaks that are shorter or surrounded by other peaks.In the feature extraction step, VMD decomposed the input audio segment into a set of IMFs. The HHT was then applied to all IMFs to create a Hilbert spectrum with a frequency resolution of 50 Hz. The Hilbert spectrum is a matrix, (H(n,f)) that contains the instantaneous energies, (h(n,f)).$$begin{aligned} H(n,f) = begin{bmatrix} h_{11} &{}dots &{} h_{1R} \ vdots &{} ddots \ h_{Q1} &{} &{} h_{QR} end{bmatrix} end{aligned}$$ (10) where r is the length of the input segment and q is the number of frequency bins in (H).The matrix (H (n,f)) is then converted from a 2-D array to a 1-D spectral representation by summing all instantaneous energy values in each frequency bin, as follows:$$begin{aligned} H(f) = sum _{n=1}^{R} H(n,f) end{aligned}$$ (11) The energy summation sequence was converted to a base-10 logarithmic scale and then smoothed by passing through a 17-point median filter and an 11-point moving average filter for the purpose of easily extracting features. All peaks and notches in the sequence whose prominence values exceeded the threshold of 0.5 were located, and their frequency values and prominence values were then stored as extracted features from the input signal (see Fig. 5).Figure 5Example of locating peak and notch frequencies and how prominent they are compared to other peaks and notches. The wave form in (a) is the smoothed energy summation sequence from the Hilbert spectrum of the Lo signal in Fig. 1. Subplot (b) is a flipped version of the energy summation sequence for the convenience of extracting notch frequencies and their prominence values. The length of the red line represents the prominence value of a peak or notch.Full size imageFor testing the effectiveness of the VMD feature extractor, a second set of features were extracted from the FFT-based power spectrum using the same input signals with the Welch’s algorithm. The FFT-based spectrum was calculated on 2048 samples with 50% overlap and a 2048-point Hanning window with 48.82 Hz frequency resolution. The power spectral density sequence was then converted to dB and went through a 21-point median filter and a 15-point moving average filter. Feature extraction followed the same strategies as in VMD feature extractor except using a prominence threshold of 2 dB.Next, the measured features, frequencies (Hz) of the peaks and notches (henceforth referred to as “measured peaks and notches”), were matched with the probability distribution functions (PDFs) of peaks and notches (henceforth referred to as “parameter peaks and notches”) from Soldevilla et al. (2008). The matching between measured and parameter peaks and notches was done in preparation of weight calculations, and it was implemented for both Gg and Lo. There are four Gaussian PDFs for parameter peaks and three for parameter notches for each species in Soldevilla et al. (2008) (Table 2). A 95% confidence interval of a Gaussian PDF was used here as a frequency range defined as 1.96 standard deviations to the left and right of its mean value. When measured peaks and notches were matched to parameter peaks and notches, only the peak or notch that fell within a 95% confidence interval were kept. Any peaks or notches outside the 95% confidence intervals were discarded.Because there are overlaps between the 95% confidence intervals of 22.4 kHz and 25.5 kHz parameter peaks of Gg and between 33.7 kHz and 37.3 kHz parameter peaks of Lo (see Table 2), it is likely that some measured peaks will fall in the overlapping areas. In this paper, the maximum a posterior (MAP) estimation41 was used to determine which PDF results in the measured peak in an overlapping area. For a measured peak that falls into an overlapping area, two parameter peaks’ PDFs are plugged in the MAP estimation equation sequentially, and then the measured peak will be matched with the PDF that maximizes the posterior probability of it given the measured peak.After the preliminary match, if more than one measured peak or notch remains in any one PDF confidence interval, the measured peak and notch with the highest prominence value is selected as the real measured peak or notch of this PDF, and the redundant ones are discarded. Finally, all remaining peak prominence values and notch prominence values were scaled to be between 0 and 1, respectively.Once peak and notch matching and selection was finished, Bayesian weights were calculated to select the most likely species. From Bayes’s rule, the posterior probability of a parameter given its measurement is proportional to the product of the likelihood function of the measurement given the parameter and the prior probability of the parameter41, as shown in Eq. (12).$$begin{aligned} P_{varvec{Theta mid Y}}(varvec{theta _i} mid varvec{y_i}) propto f_{varvec{Y mid Theta }}(varvec{y_i} mid varvec{theta _i}) P_{varvec{Theta }}(varvec{theta _i}) end{aligned}$$ (12) therefore, substitution of the posterior probability in Eq. (9) yields$$begin{aligned} w(varvec{theta _i} mid varvec{y_i}) = f_{varvec{Y mid Theta }}(varvec{y_i} mid varvec{theta _i}) *P_{varvec{Theta }}(varvec{theta _i}) * varvec{p_i} end{aligned}$$ (13) Figure 6Example of feature matching. The top plots show a set of measured peaks and notches matched with both Gg’s PDFs (a) and Lo’s PDFs (b) parameter peaks and notches like in Fig. 5 during the feature matching and selection step. Middle plots show how closely to the parameter PDFs that the measured peaks match either Gg (c) or Lo (d) and their weight calculations. The width of each PDF represents its 95% confidence interval, and the ordinate represents the weight value. Subplots (e) and (f) show the same weight calculations for notches. The final weight value is the summation of all weight values of peaks and notches matched with Gg or Lo.Full size imageWith all PDFs and a priori probabilities from Soldevilla et al. (2008), the weight value in terms of Gg and Lo given a set of measurements, (varvec{y}), was obtained by Eqs. (13) and (14)$$begin{aligned} w(Gg mid varvec{y}) = sum _{forall i} w(varvec{theta _i} mid varvec{y_i}) qquad w(Lo mid varvec{y}) = sum _{forall j} w(varvec{theta _j} mid varvec{y_j}) end{aligned}$$ (14) where (varvec{y_i}) and (varvec{y_j}) are the remaining measured peaks and notches that were matched with Gg’s PDFs and Lo’s PDFs after the matching and matching step. The feature matching and selection results and the weight calculation process are shown in Fig. 6.The last step was a comparison between weight values in terms of Gg and Lo. If (w(Lo mid varvec{y}) > w(Gg mid varvec{y})), the signal was labeled an Lo signal; otherwise, it was labeled a Gg signal. The classifier is illustrated in Fig. 7. The weight values are significant to three digits because weights are normally smaller than 1.000 and three significant digits was sufficient for comparing all calculated weight values for these audio files. In the case that the weight comparison is equal to three significant digits (even though this never happened in these 174 signals), the Bayesian VMD algorithm will automatically classify the input as a Gg signal given that the highest precision (85.91%) by the Bayesian VMD Method was achieved on Gg.Figure 7Block diagram of the Bayesian VMD Method classifier.Full size image More

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    Author Correction: Late Quaternary dynamics of Arctic biota from ancient environmental genomics

    Department of Zoology, University of Cambridge, Cambridge, UKYucheng Wang, Bianca De Sanctis, Ana Prohaska, Daniel Money & Eske WillerslevLundbeck Foundation GeoGenetics Centre, GLOBE Institute, University of Copenhagen, Copenhagen, DenmarkYucheng Wang, Mikkel Winther Pedersen, Fernando Racimo, Antonio Fernandez-Guerra, Alexandra Rouillard, Anthony H. Ruter, Hugh McColl, Nicolaj Krog Larsen, James Haile, Lasse Vinner, Thorfinn Sand Korneliussen, Jialu Cao, David J. Meltzer, Kurt H. Kjær & Eske WillerslevThe Arctic University Museum of Norway, UiT— The Arctic University of Norway, Tromsø, NorwayInger Greve Alsos, Eric Coissac, Marie Kristine Føreid Merkel, Youri Lammers & Galina GusarovaDepartment of Genetics, University of Cambridge, Cambridge, UKBianca De Sanctis & Richard DurbinUniversité Grenoble Alpes, Université Savoie Mont Blanc, CNRS, LECA, Grenoble, FranceEric CoissacCenter for Macroecology, Evolution and Climate, GLOBE Institute, University of Copenhagen, Copenhagen, DenmarkHannah Lois Owens, Carsten Rahbek & David Nogues BravoDepartment of Geosciences, UiT—The Arctic University of Norway, Tromsø, NorwayAlexandra RouillardUniversité Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Gif-sur-Yvette, FranceAdriana AlbertiGénomique Métabolique, Genoscope, Institut François Jacob, CEA, CNRS, Université Evry, Université Paris-Saclay, Evry, FranceAdriana Alberti, France Denoeud & Patrick WinckerInstitute of Earth Sciences, St Petersburg State University, St Petersburg, RussiaAnna A. Cherezova & Grigory B. FedorovArctic and Antarctic Research Institute, St Petersburg, RussiaAnna A. Cherezova & Grigory B. FedorovSchool of Geography and Environmental Science, University of Southampton, Southampton, UKMary E. EdwardsAlaska Quaternary Center, University of Alaska Fairbanks, Fairbanks, AK, USAMary E. EdwardsCentre d’Anthropobiologie et de Génomique de Toulouse, Université Paul Sabatier, Faculté de Médecine Purpan, Toulouse, FranceLudovic OrlandoNational Research University, Higher School of Economics, Moscow, RussiaThorfinn Sand KorneliussenDepartment of Geography and Environment, University of Hawaii, Honolulu, HI, USADavid W. BeilmanDepartment of Geosciences and Natural Resource Management, University of Copenhagen, Copenhagen, DenmarkAnders A. BjørkCarlsberg Research Laboratory, Copenhagen, DenmarkChristoph Dockter & Birgitte SkadhaugeCenter for Environmental Management of Military Lands, Colorado State University, Fort Collins, CO, USAJulie EsdaleFaculty of Biology, St Petersburg State University, St Petersburg, RussiaGalina GusarovaDepartment of Glaciology and Climate, Geological Survey of Denmark and Greenland, Copenhagen, DenmarkKristian K. KjeldsenDepartment of Earth Science, University of Bergen, Bergen, NorwayJan Mangerud & John Inge SvendsenBjerknes Centre for Climate Research, Bergen, NorwayJan Mangerud & John Inge SvendsenUS National Park Service, Gates of the Arctic National Park and Preserve, Fairbanks, AK, USAJeffrey T. RasicZoological Institute, , Russian Academy of Sciences, St Petersburg, RussiaAlexei TikhonovResource and Environmental Research Center, Chinese Academy of Fishery Sciences, Beijing, ChinaYingchun XingCollege of Plant Science, Jilin University, Changchun, ChinaYubin ZhangDepartment of Earth and Atmospheric Sciences, University of Alberta, Edmonton, Alberta, CanadaDuane G. FroeseCenter for Global Mountain Biodiversity, GLOBE Institute, University of Copenhagen, Copenhagen, DenmarkCarsten RahbekSchool of Environment, Earth and Ecosystem Sciences, The Open University, Milton Keynes, UKPhilip B. Holden & Neil R. EdwardsDepartment of Anthropology, Southern Methodist University, Dallas, TX, USADavid J. MeltzerDepartment of Geology, Quaternary Sciences, Lund University, Lund, SwedenPer MöllerWellcome Trust Sanger Institute, Wellcome Genome Campus, Cambridge, UKEske WillerslevMARUM, University of Bremen, Bremen, GermanyEske Willerslev More

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    Savannahs store carbon despite frequent fires

    NEWS AND VIEWS
    16 March 2022

    Savannahs store carbon despite frequent fires

    An analysis of carbon stored in the plants and soil of an African savannah suggests that atmospheric carbon dioxide concentrations — and thus global warming — might be less affected by frequent fires than we thought.

    Niall P. Hanan

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    Anthony M. Swemmer

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    Niall P. Hanan

    Niall P. Hanan is in the Jornada Basin Long-Term Ecological Research programme, Department of Plant and Environmental Sciences, New Mexico State University, Las Cruces, New Mexico 88003, USA.

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    Anthony M. Swemmer

    Anthony M. Swemmer is in the South African Environment Observation Network, Phalaborwa 1390, South Africa.

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    Savannahs burn more frequently than any other biome, and tropical savannahs alone account for 62% of the carbon dioxide emitted from fires globally1. Strategies involving fire suppression2 or the planting of trees3 in savannahs have therefore been proposed as a means of reducing CO2 emissions and increasing carbon sequestration, thus potentially contributing to the mitigation of global climate change. But it remains unclear whether these measures would make a substantial difference to the accumulation of CO2 in the atmosphere. Writing Nature, Zhou et al.4 analyse a long-term fire experiment in Kruger National Park, South Africa, and reveal that the total amount of carbon stored in the ecosystem increases more slowly than expected in the absence of fire — challenging our assumptions about how fire affects carbon storage in savannahs.

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    Nature 603, 395-396 (2022)
    doi: https://doi.org/10.1038/d41586-022-00689-0

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    Are there limits to economic growth? It’s time to call time on a 50-year argument

    EDITORIAL
    16 March 2022

    Are there limits to economic growth? It’s time to call time on a 50-year argument

    Researchers must try to resolve a dispute on the best way to use and care for Earth’s resources.

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    Lead author Donella Meadows wrote that the book The Limits to Growth “was written not to predict doom but to challenge people to find ways of living that are consistent with the laws of the planet”.Credit: Alamy

    Fifty years ago this month, the System Dynamics group at the Massachusetts Institute of Technology in Cambridge had a stark message for the world: continued economic and population growth would deplete Earth’s resources and lead to global economic collapse by 2070. This finding was from their 200-page book The Limits to Growth, one of the first modelling studies to forecast the environmental and social impacts of industrialization.For its time, this was a shocking forecast, and it did not go down well. Nature called the study “another whiff of doomsday” (see Nature 236, 47–49; 1972). It was near-heresy, even in research circles, to suggest that some of the foundations of industrial civilization — mining coal, making steel, drilling for oil and spraying crops with fertilizers — might cause lasting damage. Research leaders accepted that industry pollutes air and water, but considered such damage reversible. Those trained in a pre-computing age were also sceptical of modelling, and advocated that technology would come to the planet’s rescue. Zoologist Solly Zuckerman, a former chief scientific adviser to the UK government, said: “Whatever computers may say about the future, there is nothing in the past which gives any credence whatever to the view that human ingenuity cannot in time circumvent material human difficulties.”But the study’s lead author, Donella Meadows, and her colleagues stood firm, pointing out that ecological and economic stability would be possible if action were taken early. Limits was instrumental to the creation of the United Nations Environment Programme, also in 1972. Overall, more than 30 million copies of the book have been sold.
    The value of biodiversity is not the same as its price
    But the debates haven’t stopped. Although there’s now a consensus that human activities have irreversible environmental effects, researchers disagree on the solutions — especially if that involves curbing economic growth. That disagreement is impeding action. It’s time for researchers to end their debate. The world needs them to focus on the greater goals of stopping catastrophic environmental destruction and improving well-being.Researchers such as Johan Rockström at the Potsdam Institute for Climate Impact Research in Germany advocate that economies can grow without making the planet unliveable. They point to evidence, notably from the Nordic nations, that economies can continue to grow even as carbon emissions start to come down. This shows that what’s needed is much faster adoption of technology — such as renewable energy. A parallel research movement, known as ‘post-growth’ or ‘degrowth’, says that the world needs to abandon the idea that economies must keep growing — because growth itself is harmful. Its proponents include Kate Raworth, an economist at the University of Oxford, UK, and author of the 2017 book Doughnut Economics, which has inspired its own global movement.Economic growth is typically measured by gross domestic product (GDP). This composite index uses consumer spending, as well as business and government investment, to arrive at a figure for a country’s economic output. Governments have entire departments devoted to ensuring that GDP always points upwards. And that is a problem, say post-growth researchers: when faced with a choice between two policies (one more green than the other), governments are likely to opt for whichever is the quicker in boosting growth to bolster GDP, and that might often be the option that causes more pollution.
    G20’s US$14-trillion economic stimulus reneges on emissions pledges
    A report published last week by the World Health Organization (see go.nature.com/3j9xcpi) says that if policymakers didn’t have a “pathological obsession with GDP”, they would spend more on making health care affordable for every citizen. Health spending does not contribute to GDP in the same way that, for example, military spending does, say the authors, led by economist Mariana Mazzucato at University College London.Both communities must do more to talk to each other, instead of at each other. It won’t be easy, but appreciation for the same literature could be a starting point. After all, Limits inspired both the green-growth and post-growth communities, and both were similarly influenced by the first study on planetary boundaries (J. Rockström et al. Nature 461, 472–475; 2009), which attempted to define limits for the biophysical processes that determine Earth’s capacity for self-regulation.Opportunities for cooperation are imminent. At the end of January, the Intergovernmental Science-Policy Platform on Biodiversity and Ecosystem Services announced a big study into the causes of biodiversity loss, including the role of economic systems. More than 100 authors from 40 countries and different fields will spend two years assessing the literature. They will recommend “transformative change to the systems leading us to catastrophe”, says study co-chair, political scientist Arun Agrawal at the University of Michigan in Ann Arbor.Another opportunity is an upcoming revision of the rules for what is measured in GDP. These will be agreed by countries’ chief statisticians and organized through the UN, and are due to be finalized in 2025. For the first time, the statisticians are asking how sustainability and well-being could be more closely aligned to GDP. Both post-growth and green-growth advocates have valuable perspectives.Research can be territorial — new communities emerge sometimes because of disagreements in fields. But green-growth and post-growth scientists need to see the bigger picture. Right now, both are articulating different visions to policymakers, and there is a risk this will delay action. In 1972, there was still time to debate, and less urgency to act. Now, the world is running out of time.

    Nature 603, 361 (2022)
    doi: https://doi.org/10.1038/d41586-022-00723-1

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    Risk of colloidal and pseudo-colloidal transport of actinides in nitrate contaminated groundwater near a radioactive waste repository after bioremediation

    Characteristics of environmental samples before and after bioremediationTable 1 lists the parameters of the samples collected from the upper aquifer (12 m) at three-time points. In sample 1, before bioremediation, the content of nitrate ions reached 2517 mg/L. Against this background, in an oxidizing environment, a high content of uranium up to 1.1 mg/L and plutonium up to 0.7 Bq/L was observed. The content of organic matter did not exceed 5.9 mg/L. The suspension contained a significant amount of clay particles. Uranium in sample 1 was predominantly in dissolved form or nanoaggregates less than 5 nm in size (Fig. 1).Figure 1Percentage distribution of uranium in the filtrate during sequential filtration of samples 1 and 3. Concentrations of U in the filtrates were determined by the ICP-MS method.Full size imageIn sample 2, a year after the injection of organic matter, the content of nitrate ions reached 320 mg/L, while the values of the redox potential continued to remain in the reduction region (− 175 mV) as they were 3 months after bioremediation. The content of organic matter reached 57.5 mg/L. The uranium content dropped to 80 μg/L, and the plutonium content was below the detection limit of the device.2 years after injection (sample 3), the content of nitrate ions increased to 970 mg/L, the redox potential entered the oxidizing region and reached + 70 mV, while no significant release of uranium into solution occurred. According to the distribution scheme of uranium (Fig. 1), most of it was associated with large particles of more than 400 microns in size of clay and ferruginous nature. The plutonium content was below the sensitivity of the method. Thus, despite the fact that after a single injection of organic matter, after two years the content of nitrate ions increased markedly and the value of the redox potential returned to the oxidizing region. Nevertheless, it should be mentioned no significant remobilization of uranium and plutonium occurred. It is important to note that according to the data in Table 1, a decrease in the content of suspended matter was observed in the course of bioremediation. A discussion of the content of organic matter in the suspended matter will be carried out in the next section.Figure 2 shows electronic maps of micrographs of a filter with a maximum pore size after filtration of sample 3. It has been established that U is mainly associated with large particles (suspensions) of aluminosilicate and ferrous nature. The distribution of Al, Si, Fe and U on the surface of the filter cake was fairly uniform.Figure 2Electron micrographs of the filters with a pore size of 2400 nm surface after sample 3 filtration with elements maps (A) Al, (B) Si, (C) Fe, (D) U (SEM EDX analysis).Full size imageAlthough at low plutonium concentration it was not possible to see it by the SEM EDX method on clays, it is well known that clay minerals montmorillonite and kaolinite could have been carrier phases for Pu39. In work on the analysis of colloidal transport of radionuclides in groundwater at Yucca mountain40 uranium was found to be dominantly associated with an unidentified phase rich in Si and Fe while Pu was shown to be preferentially adsorbed onto Mn-oxides in the presence of Fe-oxides.Laboratory simulation of biogenic associative colloids formation in environmental water samples, stimulated by H2
    In a laboratory experiment with environmental samples, molecular hydrogen was used to stimulate microbial processes in order to avoid changing the content of the organic matter.Filtration studies (step-by-step filtration, Fig. 3) revealed that only 8% of organic matter in sample 1 was represented by suspended particles over 1200 nm in size. These were bacterial cells and other large particles (fulvic and humate acids, etc.). More than 50% of organic matter was in soluble form or in the form of colloidal particles up to 100 nm. In general, the distribution of organic matter in sample 3 was similar to sample 1—about 60% of organic matter was in dissolved or colloidal form and about 10% in the form of large particles.Figure 3Organic matter distribution by particle size (nm) in samples 1 and 3 before and after (B) microbial activation. Organic matter in the filtrate after each filtration step was measured using an Elementar Vario EL III CHN analyzer.Full size imageAn organic carbon content of 100 and 200 mg/L was observed in samples 1 and 2, respectively, after microbial activation by molecular hydrogen.After day 30 of incubation in sample 1 and after microbial processes, there was a noticeable increase in the content of large organic particles; their contribution reached 50%. In this case, the content of dissolved organic matter and organic particles of colloidal size decreased noticeably (their total contribution did not exceed 10% probably due to their consumption or aggregation into larger fractions). The content of organic particles with a size range of 220–450 nm had noticeably increased.In sample 3, a noticeable decrease in dissolved and colloidal organic matter was also noted; the content of organic particles of 220–100 nm and particles of 1200–400 nm increased markedly. We believe that the increase in organic particles in both samples in the range of 100–1200 nm is associated with an increase in the content of bacterial cells. Changes in the intensity of light scattering provided the most relevant information (Table 2).Table 2 The intensity of light scattering (kHz) by suspended particles of different fractions before and after day 30 of the ongoing microbial process in the stratal water (Light scattering intensity was determined by Zetasizer Nano ZS, Malvern Panalytical).Full size tableIn sample 1, before stimulation, the intensity of light scattering was at its maximum in the filtrate at 450–220 nm. In the filtrate less than 10 nm, light scattering was not detected. In filtrates larger than 450 nm and 220–50 nm, the values of the light scattering intensity were close. After microbial activation with hydrogen, a tenfold change in the intensity of light scattering was observed in the filtrate with particles larger than 2400 nm. Also, there was an almost twofold increase in filtrates with a particle size of 450–2400 nm, which is probably associated with the appearance of cells in the solution.In sample 2, before microbial activation, the maximum intensity of light scattering was observed in the filtrate with particle sizes in the range of 450–1200 nm. After microbial activation, the intensity of light scattering significantly increased in all filtrates. It is important to note that the light scattering of particles with a size characteristic of colloids (50–100 nm) increased by more than 10 times. The different behavior after hydrogen activation of two samples can probably be explained by the fact that in sample 3 the microbial community was initially more active after the injection of organic matter into the formation. In both samples, a noticeable increase in the content of coarse suspensions may indicate the agglomeration of clay suspensions by microbial polysaccharides. According to Ivanov et al.41, a similar process is observed for soil and clay particles.Laboratory simulation of the formation of biogenic associative colloids in model and environmental water samples with actinidesThe second series of experiments was carried out to evaluate the behavior of U, Np, and Pu upon activation of microbial processes. At the first stage of the laboratory simulation, a significant enlargement of large particles possibly caused by the agglomeration of natural clay and ferruginous particles due to microbial polysaccharides in natural samples was found. An important task of the second stage of the work was to assess the contribution of ferruginous and clay particles to the distribution of actinides over particles with different sizes in model solutions.When activating the microbial community in groundwater, a mixture of whey and acetate was used. However, in a laboratory simulation of this process, we decided not to use such a complex multicomponent substrate like whey. The whey contained a lot of organic suspensions and its use in this experiment would have led to even more uncertainties. A mixture of highly soluble sodium acetate and glucose substrates was added to the samples.Table 3 shows the data on the content of polysaccharides and proteins in solutions during microbial processes in samples.Table 3 Polysaccharide (A) (mg/L) and protein (B) (mg/ml) concentrations in the model solutions during incubation. Polysaccharide determination was carried out by the phenol–sulfuric acid method according to Dubois 34. Protein content was measured with the Folin phenol reagent according to Lowry 35.Full size tableNo significant increase of cells or polysaccharide content was recorded in samples with no organic matter additions. A low protein content was found in the sample NWO, which indicates that some content of cells remained in it after bioremediation. An increase in the concentration of the biomass, with peak values on day 10 and polysaccharides on day 15, was observed in all samples with additions of organic matter (O) (Table 3). The maximum accumulation of polysaccharides and protein was observed for the natural sample.On the 30th day of the experiment, there was no visible sediment in the MW sample, in the rest of the samples, there was a large amount of sediment at the bottom of the test tubes. At the same time, the solution looked almost transparent in both the MW model water sample and the MWIO sample with added iron. The average hydrodynamic radii of colloidal particles were obtained on days 3, 7, 14, 21, and 28 of the experiment (Table 4). In model water samples without added organic compounds, colloidal particles were not formed. However, by the end of the experiment, particle formation was observed. This was probably due to the transformation of colloidal matter originating from the natural water aliquot or as a result of low microbial activity.Table 4 Hydrodynamic radii of colloidal particles during the experiment, nm (The measurement accuracy was at least 2%.).Full size tableIn the presence of glucose and acetate, the emergence of the colloidal phase and a gradual increase in particle size were observed from the fifth day of incubation. The average stable hydrodynamic radii of the particles amounted to ~ 100 nm. In the presence of clay, stable colloids with the average hydrodynamic radii of 80–90 nm were formed. Stimulation of microbial processes with glucose and acetate resulted in increased particle size and partial sedimentation (samples MWO through day 20, MWIO through day 15, and NWO through day 30). After that, the sedimentation of large particles took place, and particles of smaller sizes remained in the solution.The addition of iron to the model system resulted in the formation of the particles with hydrodynamic radii of ~ 100 nm. The stimulation of the biological processes resulted in increased particle size, the formation of new particles (by day 21), and complete particle sedimentation by day 30.An important parameter used to evaluate the stability of colloidal particles in the system is the value of particles’ zeta potential. When no organic matter was added, the charge of preliminarily filtered 100–50 nm particles equaled − 29, − 26.2 mV in model water, and − 16, − 12 mV in natural water, which indicates low stability of such particles (see Table 2 Supplementary). A shift in charge of particles towards zero and positive values was observed when microbial processes were running, and this hints at the stabilization of particles in the solution.The diagrams of actinide distribution by size of colloidal particles in solutions of different nature before and after microbial stimulation on day 30 are shown in Fig. 4.Figure 4Actinide distribution by size of colloidal particles in solutions of different nature depending on the incubation time, normalized % in the filtrate. (I-before, II-after microbial stimulation on day 30). Actinides (233U, 237Np, and 239Pu) were added in the concentrations of 10–8 M/l per sample. Concentrations of 233U 239Pu were determined by liquid scintillation (Tri-Carb-3180 TR/SL liquid scintillation spectrometer) (“Perkin-Elmer,” USA).Full size imageIn the model water Pu(IV) forms true colloidal associates (up to 50%) due to deep hydrolytic polymerization. Np(V) was also partially sorbed due to slight disproportionation (by 10%). U(VI) was a stable component of soluble carbonate complexes. In the model water, increased pH and decreased Eh result in the occurrence of 99% Pu, 30% Np, and 10% U within large colloidal particles. Ultrafiltration, however, is not suitable for the assessment of the possible actinide reduction and biosorption contribution to the process of colloid formation.The microbiota and clay promote the stabilization of Pu, U, and Np in large colloidal particles. The addition of iron had no effect on actinide colloid formation, although iron caused a significant increase in neptunium colloid formation in the presence of the microbiota. This is probably due to the formation of iron-polysaccharide complexes42, which also have a high ability to chelate actinides.In Bentley More