More stories

  • in

    Visible-NIR hyperspectral classification of grass based on multivariate smooth mapping and extreme active learning approach

    Study areaGrassland herbage samples are from Shaerqin base, institute of grassland research of CAAS (Chinese Academy of Agricultural Sciences). We obtained the permission of the institution to take HSI of the grassland sample. Our work did not cause damage to grassland. Researcher Weihong Yan of the institute provided us with relevant information about grassland. The land use type in the study area is mainly grassland, which is composed of forage species, most of which are representative species of typical grassland. We take this area as an example to conduct research on grass classification. By enriching the relevant recognition technology, it can also be used as a reference for the pastures of other grasslands. The grass species Grass1 for the experiment is shown in Table 1. The official introduction of plant materials is detailed in the flora of China15.Table 1 Samples information for Grass1 dataset.Full size tableThe field hyperspectral platformWe assemble a system for collecting HSI in the field: HyperSpec©PTU-D48E HSI instrument, high-precision scanning PTZ, tripod, data analysis software Hyperspec, etc. The light source is natural light. The imaging instrument is in line scanning mode. Table 2 shows the technical parameters.Table 2 Technical parameters of hyperspectral instrument.Full size tableData collectionIn July 2021, the data was collected during the lush grass growth period. Collect data from 11:00 a.m. to 2:00 p.m. every day. At this time, it is sunny, cloudless and the wind force does not exceed level 2. So as to ensure the consistency of the acquisition time line and avoid the influence of different degrees of light on the reflectivity as far as possible. The measuring points are arranged facing the sun and the opposite direction of the shadow. We collect data from different angles of the grassland, which is based on the growth of various types of forages, and selects relatively concentrated places within the study area. Each shot is a single category of grass. The image resolution is 1166 × 1004 pixels (Fig. 1). The imaging spectrometer is fixed with scanning head when shooting. Data acquisition and transmission are executed on Hyperspec software. Then save it as a BIL file. The ENVI5.3 software was used to extract the forage spectrum to establish the dataset Grass1. Well balanced regions with a clear image, uniform spectral distribution are selected for further segmentation. The average value of spectral reflectance of grass pixels was taken as the reflectance spectrum of a single type of grass.Figure 1True color map of grass samples.Full size imageMethodologyIn Fig. 2, we present the framework of visible-NIR hyperspectral classification of grass based on multivariate smooth mapping and extreme active learning (MSM–EAL). Specifically, we first introduce the proposed MSM algorithm for global enhanced spectral reconstruction, which utilizes smooth manifold projection technology to alleviate the problems of difficult feature selection and redundant data. Then, the EAL framework is proposed to address the matter of hyperspectral labeled samples and spectral classification. In the following, each step of this method will be presented in detail.Figure 2Proposed MSM–EAL framework for grass HSI classification.Full size imageThe proposed MSM algorithmIn the process of field HSI acquisition, on the one hand, the surface distribution of grass is uneven and the plant height is different, causing certain scattering effect and coverage spectrum change. On the other hand, HSI is easy to be disturbed by external natural factors such as light, wind and shadow, resulting in a certain degree of distortion. Multiplicative scatter correction (MSC) is a scattering correction effect, which helps to eliminate the scattering effect caused by the above reasons and enhance the spectral variability. The moving window smooth spectral matrix (Nirmaf) belongs to the smooth effect, which improve the signal-to-noise ratio of the spectrum and reduce the influence of random noise16,17. Preprocessing methods are different and related to each other. We design an enhanced preprocessing multivariate smooth (MS) method that fusing MSC and smooth Nirmaf to target grass spectral signal features. In the follow-up, a model will be established to verify the validity of MS.Most of the high-dimensional spatial data have the characteristics of being embedded in a manifold body, so the manifold learning isometric feature mapping (Isomap) based on spectral theory is adopted. Isomap preserves the global geometric features of the initial data and extracts features by reconstructing the underlying smooth manifold of HSI. It is nonlinear dimensionality reduction based on linear and multidimensional scaling transformation18. Isomap has been applied in image and HSI classification19,20, but there is no report on visible-NIR hyperspectral classification of grass.In view of the above, we proposed the multivariate smooth mapping (MSM) spectral reconstruction algorithm, which can be represented as follows:$$ MSM_{z} { } = { }frac{{left( {P_{j} – b_{j} } right)left( {2n + 1} right) + n_{j} cdot mathop sum nolimits_{j = – n}^{n} C_{j} P_{k + j} }}{{n_{j} left( {2n + 1} right)}} + V_{Z} F_{Z}^{frac{1}{2}} { } $$
    (1)
    where Pj, bj, and Cj represent the raw reflectance value of spectrum j, baseline shift amount, and weight factor, respectively, k and nj represent the polynomial degree and offset, respectively. MSMz is the feature cube reconstructed to Z dimension from the spectrum calculated by 2n + 1 moving window width, V eigenvector matrix and F eigenvalue matrix.In Isomap equidistant mapping, the shortest path of edge Pi Pj needs to be solved, and the representation matrix is:$$ D_{G} = [d_{G}^{2} (P_{i} ,P_{j} )]_{i,j = 1}^{n} $$
    (2)
    where d (Pi, Pj) is the weight of the edge Pi Pj calculated from the neighborhood graph G and its side Pi Pj.The proposed EAL frameworkLabeling hyperspectral samples is expensive in terms of time and cost, at the same time, the lower spatial resolution and more bands increase the difficulty of labeling. Active learning (AL) provides an efficient labeling strategy, which only needs to label a relatively small number of samples to learn a more accurate model21. The pool-based AL selects the most informative samples according to the query strategy for limited labeling through iteration, so as to facilitate model improvement. Commonly used query strategies are uncertainty criteria, such as least confidence22, the bayesian active learning disagreement (BALD), the entropy sampling23, etc.Due to there is still an over-fitting problem, different strategies such as hybrid prediction and regularization need to be used for non-recursive datasets24. The research25 proposed that extreme gradient boosting algorithm (XGBoost) based on gradient boosting. As a classification method, XGBoost has been successfully applied in Kaggle competition and other fields. Its most important feature for visible-NIR hyperspectral classification is that can easily and directly classify according to features, and the physical interpretation of features can help understand the electronic nature behind spectral classification. XGBoost is a machine learning algorithm based tree structure that integrates multiple weak classifiers to achieve flexible and high-precision classification. It is an upgraded version of gradient boosting decision tree. The optimization process of XGBoost entailed: (1) Expanding the objective function to the second order, and finds a new objective function for the new base model to improve the calculation accuracy. (2) L2 regularization term is added to the loss function to prevent over-fitting. (3) Using blocks storage structure realize automatic parallel computing26,27. The algorithm steps are as follows:The objective function:$$ Lleft( Phi right) = mathop sum limits_{i} lleft( {y^{i} ,widehat{{y^{i} }}} right) + mathop sum limits_{k} Omega left( {f_{k} } right) $$
    (3)
    In formula (3), the first and second terms are the loss function term and the regularization term, respectively. Where,$$ Omega left( {f_{k} } right) =upgamma {text{T}} + frac{1}{2}lambda left| w right|^{2} $$
    (4)
    γ and λ are regularization parameters which are used to adjust complexity of the tree.Next, second derivative Taylor expansion of the objective function. Where (g_{i}) and (h_{i}) are the first derivative and second derivative, respectively.$$ L^{left( t right)} = mathop sum limits_{i = 1}^{n} lleft( {y_{i} ,widehat{{y_{i}^{t – 1} }} + f_{t} left( {x_{i} } right)} right) + Omega left( {f_{t} } right) $$
    (5)
    $$ g_{i} = partial_{{hat{y}_{i} (t – 1)}} lleft( {y_{i} ,widehat{{y_{i}^{t – 1} }}} right) $$
    (6)
    $$ h_{i} = partial_{{widehat{{y_{i} }}(t – 1)}}^{2} lleft( {y_{i} ,widehat{{y_{i}^{t – 1} }}} right) $$
    (7)
    $$ {text{L}}^{left( t right)} approx mathop sum limits_{i = 1}^{n} left[ {lleft( {y_{i} ,widehat{{y_{i}^{t – 1} }}} right) + g_{i} f_{i} left( {x_{i} } right) + frac{1}{2}h_{i} f_{t}^{2} left( {x_{i} } right)} right] + Omega left( {f_{t} } right) $$
    (8)
    Final objective function:$$ {hat{text{L}}}^{ i} left( q right) = – frac{1}{2}mathop sum limits_{j = 1}^{T} frac{{(mathop sum nolimits_{{i in I_{j} }} g_{i} )^{2} }}{{mathop sum nolimits_{{i in I_{j} }} h_{i} + lambda }} + gamma T $$
    (9)
    Equation (9) can be used as the fraction of tree cotyledons, and the tree structure is directly proportional to the fraction. If the result after splitting is less than the maximum value of the given parameter, the cotyledon depth stops growing24,28.AL solves the problems of limited number and high cost of grass hyperspectral labeling samples. The default model of traditional AL is logistic regression, which is mostly studied on the ideal public dataset. However, the actual data has more uncertain noise, which still poses a certain challenge to AL. Consequently, we propose the extreme active learning (EAL) framework to minimize the classification cost of visible-NIR hyperspectral. The framework replaces the logistic regression model with XGBoost. Taking advantage of AL, XGBoost can improve performance with less training marker samples. By jointing of XGBoost and AL, EAL provides significantly better results than AL in field Grassl dataset recognition. Additionally, based on the characteristics of XGBoost, EAL more intuitively enhances the physical essence behind spectral classification than AL. Algorithm 1 summarizes the workflow of EAL framework.Random forest (RF) and decision tree (DT) were used to compare with EAL. RF and DT are frequently used in the field of grassland remote sensing9,29. Furthermore, RF, DT and XGBoost have the same point is that are learning algorithms based on tree structure. DT determines the direction by judging the conditions of the decision node12. RF is an integrated learning of multiple decision trees30. More

  • in

    Mapping phyllosphere microbiota interactions in planta to establish genotype–phenotype relationships

    Flemming, H. C. & Wuertz, S. Bacteria and archaea on Earth and their abundance in biofilms. Nat. Rev. Microbiol. 17, 247–260 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Bulgarelli, D., Schlaeppi, K., Spaepen, S., Ver Loren van Themaat, E. & Schulze-Lefert, P. Structure and functions of the bacterial microbiota of plants. Annu Rev. Plant Biol. 64, 807–838 (2013).CAS 
    PubMed 
    Article 

    Google Scholar 
    Turnbaugh, P. J. et al. The human microbiome project. Nature 449, 804–810 (2007).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Venturelli, O. S. et al. Deciphering microbial interactions in synthetic human gut microbiome communities. Mol. Syst. Biol. 14, e8157 (2018).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Foster, K. R. & Bell, T. Competition, not cooperation, dominates interactions among culturable microbial species. Curr. Biol. 22, 1845–1850 (2012).CAS 
    PubMed 
    Article 

    Google Scholar 
    Helfrich, E. J. N. et al. Bipartite interactions, antibiotic production and biosynthetic potential of the Arabidopsis leaf microbiome. Nat. Microbiol. 3, 909–919 (2018).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Coyte, K. Z. & Rakoff-Nahoum, S. Understanding competition and cooperation within the mammalian gut microbiome. Curr. Biol. 29, 538–544 (2019).Article 
    CAS 

    Google Scholar 
    Turner, T. R. et al. Comparative metatranscriptomics reveals kingdom level changes in the rhizosphere microbiome of plants. ISME J. 7, 2248–2258 (2013).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Trivedi, P., Leach, J. E., Tringe, S. G., Sa, T. & Singh, B. K. Plant–microbiome interactions: from community assembly to plant health. Nat. Rev. Microbiol. 18, 607–621 (2020).CAS 
    PubMed 
    Article 

    Google Scholar 
    Müller, D. B., Vogel, C., Bai, Y. & Vorholt, J. A. The plant microbiota: systems-level insights and perspectives. Annu. Rev. Genet. 50, 211–234 (2016).PubMed 
    Article 
    CAS 

    Google Scholar 
    Lugtenberg, B. & Kamilova, F. Plant-growth-promoting Rhizobacteria. Annu. Rev. Microbiol. 63, 541–556 (2009).CAS 
    PubMed 
    Article 

    Google Scholar 
    Berendsen, R. L., Pieterse, C. M. J. & Bakker, P. A. H. M. The rhizosphere microbiome and plant health. Trends Plant Sci. 17, 478–486 (2012).CAS 
    PubMed 
    Article 

    Google Scholar 
    Innerebner, G., Knief, C. & Vorholt, J. A. Protection of Arabidopsis thaliana against leaf-pathogenic Pseudomonas syringae by Sphingomonas strains in a controlled model system. Appl. Environ. Microbiol. 77, 3202–3210 (2011).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Shekhawat, K. et al. Root endophyte induced plant thermotolerance by constitutive chromatin modification at heat stress memory gene loci. EMBO Rep. 22, e51049 (2021).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Vorholt, J. A. Microbial life in the phyllosphere. Nat. Rev. Microbiol. 10, 828–840 (2012).CAS 
    PubMed 
    Article 

    Google Scholar 
    Bodenhausen, N., Bortfeld-Miller, M., Ackermann, M. & Vorholt, J. A. A synthetic community approach reveals plant genotypes affecting the phyllosphere microbiota. PLoS Genet. 10, e1004283 (2014).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Reisberg, E. E., Hildebrandt, U., Riederer, M. & Hentschel, U. Distinct phyllosphere bacterial communities on Arabidopsis wax mutant leaves. PLoS ONE 8, e78613 (2013).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Kniskern, J. M., Traw, M. B. & Bergelson, J. Salicylic acid and jasmonic acid signaling defense pathways reduce natural bacterial diversity on Arabidopsis thaliana. Mol. Plant Microbe Interact. 20, 1512–1522 (2007).CAS 
    PubMed 
    Article 

    Google Scholar 
    Pfeilmeier, S. et al. The plant NADPH oxidase RBOHD is required for microbiota homeostasis in leaves. Nat. Microbiol. 6, 852–864 (2021).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Chen, T. et al. A plant genetic network for preventing dysbiosis in the phyllosphere. Nature 580, 653–657 (2020).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Hassani, M. A., Duran, P. & Hacquard, S. Microbial interactions within the plant holobiont. Microbiome 6, 58 (2018).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Lidicker, W. Z. Clarification of interactions in ecological systems. Bioscience 29, 475–477 (1979).Article 

    Google Scholar 
    Schlechter, R. O., Miebach, M. & Remus-Emsermann, M. N. P. Driving factors of epiphytic bacterial communities: a review. J. Adv. Res. 19, 57–65 (2019).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Faust, K. & Raes, J. Microbial interactions: from networks to models. Nat. Rev. Microbiol. 10, 538–550 (2012).CAS 
    PubMed 
    Article 

    Google Scholar 
    Grosskopf, T. & Soyer, O. S. Synthetic microbial communities. Curr. Opin. Microbiol. 18, 72–77 (2014).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Blair, P. M. et al. Exploration of the biosynthetic potential of the Populus microbiome. mSystems 3, e00045-00018 (2018).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Suda, W., Nagasaki, A. & Shishido, M. Powdery mildew-infection changes bacterial community composition in the phyllosphere. Microbes Environ. 24, 217–223 (2009).PubMed 
    Article 

    Google Scholar 
    Manching, H. C., Balint-Kurti, P. J. & Stapleton, A. E. Southern leaf blight disease severity is correlated with decreased maize leaf epiphytic bacterial species richness and the phyllosphere bacterial diversity decline is enhanced by nitrogen fertilization. Front. Plant Sci. 5, 403 (2014).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Agler, M. T. et al. Microbial hub taxa link host and abiotic factors to plant microbiome variation. PLoS Biol. 14, 100235 (2016).Article 
    CAS 

    Google Scholar 
    Layeghifard, M., Hwang, D. M. & Guttman, D. S. Disentangling interactions in the microbiome: a network perspective. Trends Microbiol. 25, 217–228 (2017).CAS 
    PubMed 
    Article 

    Google Scholar 
    Faust, K. et al. Microbial co-occurrence relationships in the human microbiome. PLoS Comput. Biol. 8, e1002606 (2012).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Carr, A., Diener, C., Baliga, N. S. & Gibbons, S. M. Use and abuse of correlation analyses in microbial ecology. ISME J. 13, 2647–2655 (2019).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Vorholt, J. A., Vogel, C., Carlström, C. I. & Müller, D. B. Establishing causality: opportunities of synthetic communities for plant microbiome research. Cell Host Microbe 22, 142–155 (2017).CAS 
    PubMed 
    Article 

    Google Scholar 
    Bai, Y. et al. Functional overlap of the Arabidopsis leaf and root microbiota. Nature 528, 364–369 (2015).CAS 
    PubMed 
    Article 

    Google Scholar 
    Knief, C., Frances, L. & Vorholt, J. A. Competitiveness of diverse Methylobacterium strains in the phyllosphere of Arabidopsis thaliana and identification of representative models, including M. extorquens PA1. Microb. Ecol. 60, 440–452 (2010).PubMed 
    Article 

    Google Scholar 
    Fan, J., Crooks, C. & Lamb, C. High-throughput quantitative luminescence assay of the growth in planta of Pseudomonas syringae chromosomally tagged with Photorhabdus luminescens luxCDABE. Plant J. 53, 393–399 (2008).CAS 
    PubMed 
    Article 

    Google Scholar 
    Carlström, C. I. et al. Synthetic microbiota reveal priority effects and keystone strains in the Arabidopsis phyllosphere. Nat. Ecol. Evol. 3, 1445–1454 (2019).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Vogel, C. M., Potthoff, D. M., Schäfer, M., Barandun, N. & Vorholt, J. A. Protective role of the Arabidopsis leaf microbiota against a bacterial pathogen. Nat. Microbiol. 6, 1537–1548 (2021).CAS 
    PubMed 
    Article 

    Google Scholar 
    Chen, I.-M. A. et al. The IMG/M data management and analysis system v.6.0: new tools and advanced capabilities. Nucleic Acids Res. 49, 751–763 (2020).Article 
    CAS 

    Google Scholar 
    Ortiz, A., Vega, N. M., Ratzke, C. & Gore, J. Interspecies bacterial competition regulates community assembly in the C. elegans intestine. ISME J. 15, 2131–2145 (2021).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Goberna, M. & Verdú, M. Predicting microbial traits with phylogenies. ISME J. 10, 959–967 (2016).PubMed 
    Article 

    Google Scholar 
    Webb, C. O., Ackerly, D. D., McPeek, M. A. & Donoghue, M. J. Phylogenies and community ecology. Annu. Rev. Ecol. Syst. 33, 475–505 (2002).Article 

    Google Scholar 
    Cahill, J. F., Kembel, S. W., Lamb, E. G. & Keddy, P. A. Does phylogenetic relatedness influence the strength of competition among vascular plants? Perspect. Plant Ecol. 10, 41–50 (2008).Article 

    Google Scholar 
    Maherali, H. & Klironomos, J. N. Influence of phylogeny on fungal community assembly and ecosystem functioning. Science 316, 1746–1748 (2007).CAS 
    PubMed 
    Article 

    Google Scholar 
    Duncan, R. P. & Williams, P. A. Ecology – Darwin’s naturalization hypothesis challenged. Nature 417, 608–609 (2002).CAS 
    PubMed 
    Article 

    Google Scholar 
    Slingsby, J. A. & Verboom, G. A. Phylogenetic relatedness limits co-occurrence at fine spatial scales: evidence from the schoenoid sedges (Cyperaceae: Schoeneae) of the Cape Floristic Region, South Africa. Am. Nat. 168, 14–27 (2006).PubMed 
    Article 

    Google Scholar 
    Mayfield, M. M. & Levine, J. M. Opposing effects of competitive exclusion on the phylogenetic structure of communities. Ecol. Lett. 13, 1085–1093 (2010).PubMed 
    Article 

    Google Scholar 
    Teixeira, P. J. P. L., Colaianni, N. R., Fitzpatrick, C. R. & Dangl, J. L. Beyond pathogens: microbiota interactions with the plant immune system. Curr. Opin. Microbiol. 49, 7–17 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Maier, B. A. et al. A general non-self response as part of plant immunity. Nat. Plants 7, 696–705 (2021).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Friedman, J., Higgins, L. M. & Gore, J. Community structure follows simple assembly rules in microbial microcosms. Nat. Ecol. Evol. 1, 0109 (2017).Article 

    Google Scholar 
    Kehe, J. et al. Positive interactions are common among culturable bacteria. Sci. Adv. 7, eabi7159 (2021).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Lindow, S. E. & Brandl, M. T. Microbiology of the phyllosphere. Appl. Environ. Microbiol. 69, 1875–1883 (2003).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Remus-Emsermann, M. N. P. et al. Spatial distribution analyses of natural phyllosphere-colonizing bacteria on Arabidopsis thaliana revealed by fluorescence in situ hybridization. Environ. Microbiol. 16, 2329–2340 (2014).CAS 
    PubMed 
    Article 

    Google Scholar 
    Billick, I. & Case, T. J. Higher-order interactions in ecological communities – what are they and how can they be detected. Ecology 75, 1529–1543 (1994).Article 

    Google Scholar 
    Grilli, J., Barabas, G., Michalska-Smith, M. J. & Allesina, S. Higher-order interactions stabilize dynamics in competitive network models. Nature 548, 210–213 (2017).CAS 
    PubMed 
    Article 

    Google Scholar 
    Levine, J. M., Bascompte, J., Adler, P. B. & Allesina, S. Beyond pairwise mechanisms of species coexistence in complex communities. Nature 546, 56–64 (2017).CAS 
    PubMed 
    Article 

    Google Scholar 
    Sundarraman, D. et al. Higher-order interactions dampen pairwise competition in the zebrafish gut microbiome. mBio 11, e01667-20 (2020).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Morris, C. in Encyclopedia for Life Sciences (National Publishing Group, 2002).Raaijmakers, J. M. & Mazzola, M. Diversity and natural functions of antibiotics produced by beneficial and plant pathogenic bacteria. Annu. Rev. Phytopathol. 50, 403–424 (2012).CAS 
    PubMed 
    Article 

    Google Scholar 
    Iversen, O. J. & Grov, A. Studies on lysostaphin – separation and characterization of 3 enzymes. Eur. J. Biochem. 38, 293–300 (1973).CAS 
    PubMed 
    Article 

    Google Scholar 
    Recsei, P. A., Gruss, A. D. & Novick, R. P. Cloning, sequence, and expression of the lysostaphin gene from Staphylococcus simulans. Proc. Natl Acad. Sci. USA 84, 1127–1131 (1987).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Kessler, E., Safrin, M., Abrams, W. R., Rosenbloom, J. & Ohman, D. E. Inhibitors and specificity of Pseudomonas aeruginosa LasA. J. Biol. Chem. 272, 9884–9889 (1997).CAS 
    PubMed 
    Article 

    Google Scholar 
    Trayer, H. R. & Buckley, C. E. Molecular properties of lysostaphin, a bacteriolytic agent specific for Staphylococcus aureus. J. Biol. Chem. 245, 4842–4846 (1970).CAS 
    PubMed 
    Article 

    Google Scholar 
    Heymer, B. & Schmidt, W. C. Purification and characterization of a Streptomyces albus endo-N-acetylmuramidase lytic for group A and other beta hemolytic streptococci. Microbios 12, 51–66 (1975).CAS 
    PubMed 

    Google Scholar 
    Vollmer, W., Joris, B., Charlier, P. & Foster, S. Bacterial peptidoglycan (murein) hydrolases. FEMS Microbiol. Rev. 32, 259–286 (2008).CAS 
    PubMed 
    Article 

    Google Scholar 
    Peyraud, R. et al. Demonstration of the ethylmalonyl-CoA pathway by using C-13 metabolomics. Proc. Natl Acad. Sci. USA 106, 4846–4851 (2009).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Schlesier, B., Breton, F. & Mock, H. P. A hydroponic culture system for growing Arabidopsis thaliana plantlets under sterile conditions. Plant Mol. Biol. Rep. 21, 449–456 (2003).CAS 
    Article 

    Google Scholar 
    Paradis, E. & Schliep, K. ape 5.0: an environment for modern phylogenetics and evolutionary analyses in R. Bioinformatics 35, 526–528 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Revell, L. J. phytools: an R package for phylogenetic comparative biology (and other things). Methods Ecol. Evol. 3, 217–223 (2012).Article 

    Google Scholar 
    Edgar, R. C. Search and clustering orders of magnitude faster than BLAST. Bioinformatics 26, 2460–2461 (2010).CAS 
    PubMed 
    Article 

    Google Scholar 
    Pruesse, E. et al. SILVA: a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB. Nucleic Acids Res. 35, 7188–7196 (2007).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Integrated Development Environment for R (R Studio, 2020).R: A Language and Environment for Statistical Computing (R Foundation for Statistical Computing, 2021).Love, M. I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 15, 550 (2014).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Oksanen, J. et al. vegan: Community Ecology Package. R package v. 2.5-7 (2020).Armenteros, J. J. A. et al. SignalP 5.0 improves signal peptide predictions using deep neural networks. Nat. Biotechnol. 37, 420–423 (2019).Article 
    CAS 

    Google Scholar 
    Gasteiger, E. et al. in The Proteomics Protocols Handbook 571–607 (ed Walker, J. M.) (Humana Press, 2005).Schindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).CAS 
    PubMed 
    Article 

    Google Scholar 
    Bushnell, B. BBMap short read aligner, and other bioinformatic tools (SourceForge, version 38.87); https://sourceforge.net/projects/bbmapDeatherage, D. E. & Barrick, J. E. Identification of mutations in laboratory-evolved microbes from next-generation sequencing data using breseq. Methods Mol. Biol. 1151, 165–188 (2014).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Kolmogorov, M., Yuan, J., Lin, Y. & Pevzner, P. A. Assembly of long, error-prone reads using repeat graphs. Nat. Biotechnol. 37, 540–546 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Walker, B. J. et al. Pilon: an integrated tool for comprehensive microbial variant detection and genome assembly improvement. PLoS ONE 9, e112963 (2014).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Seemann, T. Prokka: rapid prokaryotic genome annotation. Bioinformatics 30, 2068–2069 (2014).CAS 
    PubMed 
    Article 

    Google Scholar  More

  • in

    A noble extended stochastic logistic model for cell proliferation with density-dependent parameters

    Stability analysis of the deterministic modelSolving (left( x(t) times left( r_{p}x(t)^{(alpha )}left( 1-big (frac{x(t)}{K}big )^{beta }right) – nx(t)^{(delta )} right) right) =0), we obtain two stable and one unstable equilibrium points for the model. One stable equilibrium is trivial, i.e., (x(t)=0), another stable equilibrium point being the non-zero satisfying (left( r_{p}x(t)^{(alpha )}left( 1-big (frac{x(t)}{K}big )^{beta }right) – nx(t)^{(delta )} right) =0). Figure 1a shows three different equilibrium points of the model. In addition to the equilibrium, the model has two inflection points (Fig. 1a). At these inflection points the absolute growth rates are minimum and maximum. The density vs relative proliferation rate (RPR) profile of the model shows that the model can attain negative RPR for a positive cell density, suggesting that the model can portray the Allee phenomenon (Fig. 1b). Figure 1c,d portray the proliferation and decay phases, respectively through the model.Figure 1Growth dynamics of the proposed model: (a) Absolute proliferation rate (APR) profile considering (r_{p}=0.13), (K=1.43), (n=0.0095), (alpha =1.15), (beta =0.99) and (delta =0.2); (b) RPR profiles for different n and other same constant model parameters; (c) Cell population survive for (r_{p}=0.13), (K=1.43), (n=0.0095), (alpha =1.15), (beta =0.99) and (delta =0.2) with the initial cell density 0.1; (d) The population goes to extinction for the initial cell density 0.06 with the same constant parameters.Full size imageThe solution of the deterministic model finally provides two theorems.
    Theorem 1

    (x^{*}approx K -Kleft( frac{Big (beta r_{p}K^{alpha }+n delta K^{delta }Big )-sqrt{Big (beta r_{p}K^{alpha }+n delta K^{delta }Big )^{2}-2 left( 2 alpha beta r_{p}K^{alpha } +beta (beta -1)r_{p}K^{alpha }+delta (delta -1)nK^{delta } right) nK^{delta }}}{left( 2 alpha beta r_{p}K^{alpha } +beta (beta -1)r_{p}K^{alpha }+delta (delta -1)nK^{delta } right) }right)) is the conditional MSSCD for the intercellular-interaction-induced proliferative cells. The conditional threshold density for cell-proliferation upon interaction is (x^{*}=K -Kleft( frac{Big (beta r_{p}K^{alpha }+n delta K^{delta }Big )+sqrt{Big (beta r_{p}K^{alpha }+n delta K^{delta }Big )^{2}-2 left( 2 alpha beta r_{p}K^{alpha } +beta (beta -1)r_{p}K^{alpha }+delta (delta -1)nK^{delta } right) nK^{delta }}}{left( 2 alpha beta r_{p}K^{alpha } +beta (beta -1)r_{p}K^{alpha }+delta (delta -1)nK^{delta } right) }right)) (proof is in the supplementary information).
    Allee and cooperation models are the only extended logistic law other than our model to provide a threshold population size for growth or proliferation. Our proposed model is superior to the Allee and cooperation model as it can detect the conditional threshold cell density for proliferation and regulate the density by its different parameters. For example, One may reduce the conditional threshold density by either regulating the interaction between growth-inhibiting molecules and cells ((delta)) or reducing the inhibiting molecule concentration (n).The conditional MSSCD from Theorem 1 is lower than the carrying capacity of the conventional logistic model due to growth-inhibiting molecules; it provides the expected cell density during culture in a given environment. Theorem 1 also states the set of parameters to control the cell proliferation and get the desired density during such cell cultures. A further question arises knowing this set of parameters: which one of the parameters in the expression is crucial in terms of application purpose? Since the (r_{p}) is the constant proliferation rate for a given cell line, controlling the conditional MSSCD is not possible through (r_{p}). We simulate the distribution of conditional MSSCD for other parametric planes to answer this question. For this, we use the parameter values obtained from the data.

    Theorem 2

    The RPR is maximum at the cell density (x^{*}= K-Kleft( frac{r_{p}beta K^{alpha -1}+ndelta K^{delta -1}}{2r_{p}alpha beta K^{alpha -1}+r_{p}beta (beta -1)K^{alpha -1}+ndelta (delta -1)K^{delta -1}}right)) for the concave downward profile under the condition (r_{p}alpha (alpha -1){x^{*}}{}^{(alpha -2)}-frac{r_{p}}{K^{beta }}(alpha +beta )(alpha +beta -1){x^{*}}{}^{(alpha +beta -2)}-ndelta (delta -1){x^{*}}{}^{(delta -2)}n) (see the supplementary information). The cell population sustain with any positive initial cell density x(t) and try to stabilize at (x(t)= K(1-frac{n}{r_{p}})^frac{1}{beta }). Therefore, bimodality vanishes and unimodality is observed for the case (alpha =delta) (r_{p} >n). The RPR profile will be concave downward always with the maximum RPR value is at the inflection point (x(t)= K(frac{(r_{p}-n)alpha }{r_{p}(alpha +beta )})^frac{1}{beta }). The deterministic potential function in this case is (U(x)=-Big [(r_{p}-n)frac{x^{(alpha +2)}}{(alpha +2)}-frac{r_{p}}{K^{beta }}frac{x^{(alpha +beta +2)}}{(alpha +beta +2)} Big ]). The minima of this effective potential function will be at (x(t)= K(1-frac{n}{r_{p}})^frac{1}{beta }) which is the maximum stable cell density for (r_{p} >n).
    Parameter estimationThe density-RPR and time-density fitting to the scratch assay datasets show a lower RSS for our model than the logistic one for each of the three seeding conditions. The estimated parameters from the RPR fitting through the grid-search are in Table 2. Although the RSS for the RPR fitting of the seeding 2 is very low, the data itself is too scattered in both the upper and lower range for the small cell density. Therefore, there is a chance that regardless of the low RSS value, the fitting for seeding 2 may not reflect the actual estimates of the parameters with the bias in the data set (Fig. 2b). Nevertheless, the density-RPR fittings to the other two seeding density datasets do not suffer from bias (Fig. 2a,c).Table 2 Estimated model parameters from density-RPR fitting of our model.Full size table
    Figure 2Our proposed model best fitted the cell density-RPR datasets for all of the seeding conditions generated through the grid-search method.Full size image
    Jin et al.1 suggested that their two phase logistic model may share similarities with the Allee effect. However, they did not fit the Allee model stating seeding 2 and 3 were large enough seeding densities. We calculated the conditional threshold density, conditional MSSCD, density at the minimum and maximum RPR for the model from our estimated parameters (Table 3). The conditional threshold cell density calculated from our estimated parameters confirms that the smallest initial seeding density of the dataset was greater than the conditional threshold cell density.Table 3 Calculated cell densities from estimated parameters from our model fitting.Full size tableFigure 3 compares the portrayal of the data through our model with the fitting by Jin et al.1. The blue dashed line is the time-series fitting of the proposed model, and the red-colored line is the time-series fitting of the logistic model to the scratch assay data sets in the Fig. 3. The carrying capacity values are unexpectedly very high in the logistic fit, keeping the model near the exponential phase for the entire dataset. Thus the overall and two phase logistic fits are unrealistic compared to the highest cell density observed in the assay. Also, logistic fitting of the RPR profiles to the data after 18 h does not capture the whole scenario. The green solid and the violet dashed line represent the logistic time-density fit after and before 18 h density profiles respectively. The orange-colored lines in the Fig. 3 are the expected population density as per estimated parameters from the RPR fitting after 18 h data sets. Table 4 enlists all parameters for a comparison between logistic and our model fitting.Figure 3Time series solution of the proposed model and logistic law with comparative RSS for all three seeding conditions.Full size imageTable 4 Logistic model fitting with the Jin et al.1 estimates used in Fig. 3 with the specific colors.Full size tableTrends in cell densities under deterministic set upThe (r_{p}) is fixed for a cell line among all the determining parameters of the conditional MSSCD. n and K vary together with the culture media, flask, and environmental setup. On the other hand, the (alpha), (beta), and (delta) vary together with intercellular-interactions and cellular-interaction with growth-inhibitory molecules, which depend on the medium’s initial cell density per well and fluidity. We observe that the distribution of the conditional MSSCD depends more on the K than the n. There is a chance of overproliferation in the deterministic setup under low n but high K. The cells may die under high n. The cell density at maximum RPR also depends more on K than n (Fig. 4). So the cells should be cultured in the larger flask to achieve maximum proliferativeness.Figure 4The distribution of conditional MSSCD and cell density at maximum RPR in n-K parametric plane.Full size imageThe conditional MSSCD depends more on (beta) than (alpha) (Fig. 5a). The cells may tend to overproliferate under both high (alpha) and (beta). The conditional MSSCD does not exist for a high (delta) and low (beta) depending more on (delta) than (beta). The cells may overproliferate only under a high (beta) and low (delta) (Fig. 5b). The conditional MSSCD also depends more on (delta) than (alpha) showing mostly underproliferation of cells in the (delta ~-alpha) parametric plane. Therefore, the proliferation can be controlled via regulating the interaction between the growth-inhibitory molecules and cells followed by density-regulation through contact-inhibition and cell-cell cooperation (Fig. 5c).Figure 5The distribution of the conditional MSSCD in parametric plane of regulators in the growth law: (a) dependence of the conditional MSSCD on (alpha) and (beta) parameters; (b) dependence of the conditional MSSCD on (delta) and (beta) parameters; (c) dependence of the conditional MSSCD on (alpha) and (delta) parameters.Full size imageThe new cell fitness measure, i.e. cell density at maximum RPR depends more on the (alpha) than the (beta) (Fig. 6a). The cells achieve maximum RPR at a great cell density under the high value of these two parameters. Figure 6b,c suggest that cell density depends only a little on the (delta) under high (alpha) and (beta). Under the low value of these two regulators, a high (delta) always reduces the cell density attaining the maximum RPR, resulting a poor cell-fitness.Figure 6The distribution of cell density at maximum RPR in parametric plane of regulators in the growth law: (a) dependence on (alpha) and (beta) parameters; (b) dependence on (alpha) and (delta) parameters; (c) dependence on (delta) and (beta) parameters.Full size imageStochastic model analysisOur proposed stochastic model (3) can be compared with the general stratonovich stochastic differential equation (frac{dx}{dt}=f(x)+g_{1}(x)epsilon (t)+g_{2}(x)Gamma (t)). Comparing it with our proposed stochastic model we obtain (g_{1}(x)=-x^{delta +1}) and (g_{2}(x)=1). Using the help of47, we get noise induced drift (A(x)=r_{p}x^{alpha +1}left( 1-Big (frac{x}{K}Big )^{beta } right) -nx^{(delta +1)}+D(delta +1)x^{(2delta +1)}-lambda sqrt{DQ}(delta +1)x^{delta }) and noise induced diffusion coefficient (B(x)=Dx^{(2delta +2)}-2lambda sqrt{DQ}x^{(delta +1)}+Q). The cell density at long run can be obtained from the steady state probability density function (SSPDF). The analytical expression of the SSPDF is obtained from the Fokker-Planck equation. The Fokker-Planck equation is (frac{partial P(x, t)}{partial t} =- frac{partial big [ A(x) P(x, t)big ]}{partial x}+ frac{partial ^{2} big [B(x) P(x, t)big ]}{partial x^{2}}), where P(x,t) is the probability density function of the cell population at the time point t. Solving the Fokker-Planck equation we get the SSPDF as (P_{st} (x)= frac{N^{prime }}{B(x)} exp left( int _{x} frac{A(x^{prime })}{B(x^{prime })} dx^{prime }right)) with the normalizing constant (N^{prime }). The value of (N^{prime }) can be obtained from (int _{0}^{infty } P_{st} (x)dx=1).This SSPDF (P_{st} (x)) helps to understand the validity of the proposed stochastic model. Since the number of the data points is too low to fit the stochastic model to the data directly, validation of the stochastic model is challenging in this case. The dataset we used is a time series with 15 data points with three replicates only. An experiment must have many replicates to have a sample with a large sample size so that the SSPDF of cell densities obtained from theoretical findings can be validated with the real observation of cell densities at the steady state. Such datasets with many replicates are rare.So, we generate 2000 sample paths with the help of numerical simulation based on stochastic model 3. We use the parameter values estimated from the fittings of the deterministic model to the seeding condition 1, and we consider some particular values for the two noise intensities and correlation strength ((lambda)) to get a simulated dataset. To achieve the stationary state, we consider sufficiently large time points, and the cell densities at the final time point are used as the data set for the stationary state. We compare the frequency density of cell densities at steady-state of a simulated dataset of 2000 sample paths with the SSPDF obtained from the analytical solution. This comparison shows that the cell density distribution at the steady state matches the steady state probability density function obtained analytically (Fig. 7).In addition, we illustrated the time series generated with the help of stochastic model 3 through numerical technique (Fig. 8). We have plotted the time series data thus obtained for each of the three seeding conditions and in the same figure we also plotted the observed cell densities. The red dots (o) represent the original/experimental dataset of Jin et al.1. The blue dots ((*)) represent the simulated dataset obtained from the stochastic model. This Fig. 8 clarifies our claim that the proposed stochastic model is in good agreement with the actual observation.Figure 7The histogram shows the distribution of cell densities at steady state under additive and multiplicative noises. The blue curve is the SSPDF. The function SSPDF and the distribution of cell densities matches to each other.Full size imageFigure 8The red dots (o) in each sub-figures represent the experimental data of Jin et al.1. The blue dots ((*)) are obtained from the stochastic model (3) considering: (a) The seeding 1 estimated model parameters with (D= 0.002), (Q= 0.06) and (lambda = 0.4). (b) The seeding 2 estimated model parameters with (D= 0.01), (Q= 0.15) and (lambda = 0.6). (c) The seeding 3 estimated model parameters with (D= 0.002), (Q= 0.2) and (lambda = 0.4).Full size imageFigures 7 and 8 suggest that the stochastic model is valid. So the model can be further analyzed to meet the first objective. Differentiating (P_{st} (x)), we obtain (frac{dP_{st} (x)}{dx}=frac{N^{prime }}{[B(x)]^2} exp left( int frac{A(x)}{B(x)}dx right) left( A(x)-frac{dB(x)}{dx} right)) and (frac{d^{2}P_{st} (x)}{dx^{2}}= frac{N^{prime }}{[B(x)]^{2}}exp left( int frac{A(x)}{B(x)}dx right) left( frac{dA(x)}{dx}-frac{d^{2}B(x)}{dx^{2}} right) +frac{N^{prime }}{[B(x)]^{2}} left( A(x)-frac{dB(x)}{dx} right) exp left( int frac{A(x)}{B(x)}dx right) frac{A(x)}{B(x)}-frac{2}{[B(x)]^3}N^{prime } exp left( int frac{A(x)}{B(x)}dx right) left( A(x)-frac{dB(x)}{dx} right) frac{dB(x)}{dx}). At the extrema of the SSPDF, we must have (frac{dP_{st} (x)}{dx}=0) i.e. (left( A(x)-frac{dB(x)}{dx} right) =0).

    Theorem 3

    (x^{*}approx K-K left( frac{nK^{delta +1}+D(delta +1) K^{2delta +1}-lambda sqrt{DQ}(delta +1)K^{delta }}{beta r K^{alpha +1}+n(delta +1) K^{(delta +1)}+D(delta +1) (2delta +1)K^{(2delta +1)}-lambda sqrt{DQ}delta (delta +1)K^{delta }} right)) is the conditional MSSCD due to the correlated additive and multiplicative noises under the condition (r_{p}(alpha +1)x^{*}{}^{alpha }-frac{r_{p}}{K^{beta }}(alpha +beta +1)x^{*}{}^{(alpha +beta )} -n(delta +1)x^{*}{}^{delta }-D(delta +1)(2delta +1)x^{*}{}^{(2delta )}+lambda sqrt{Dalpha }delta (delta +1)x^{*}{}^{(delta -1)} < 0) (proof is in the supplementary information). Figure 9 visualizes the effect of noise strength and correlation strength on the conditional MSSCD. The conditional MSSCD increases with the additive noise strength (Q) and decreases with the multiplicative noise strength (D) when the other model parameters are fixed (Fig. 9a). There is a high chance of overproliferation for a low D and a high Q (Fig. 9a). Again, there is a high chance of extinction for the low Q and high D. The conditional MSSCD depends more on D than (lambda) (Fig. 9b), and more on (lambda) than Q (Fig. 9c). The conditional MSSCD increases with (lambda) and Q; there is a high chance of overproliferation for high (lambda) and Q. The extinction risk of cells from the culture increases with low (lambda) and Q.Figure 9The change in the conditional MSSCD value for different noise strengths and correlation strength using the parameters estimated for seeding 1: (a) the conditional MSSCD values in (D-Q) noise strength plane with highest correlation ((lambda =1)); (b) the conditional MSSCD values in (D-lambda) noise plane with (Q=0.01); (c) the conditional MSSCD values in (Q-lambda) noise plane with (D=0.01).Full size imageDue to the difficulty and complicated expression of the analytical expression of the SSPDF, we use numerical simulation to study the steady-state behavior in the long run under correlated noises. We draw a histogram of the cell densities based on 500 normal sample paths at the final time points. We use seeding 1 fitting estimates as the initial parameter values for this simulation. The cell population is stable and steady at either 0 cell density or at the conditional MSSCD. The distribution is symmetric around the conditional MSSCD for (lambda =1) (Fig. 10a). There is a loss in the symmetry for the decreasing (lambda). For (lambda =0.5), there is a mode at the zero states with another mode at conditional MSSCD (Fig. 10b). The histogram shows a bi-modality for low values of (lambda). The mode at the zero state is highest for (lambda =0) (Fig. 10c). Therefore, the extinction chance increases for zero noise correlation between the additive and the multiplicative noises.Figure 10Distribution of cell density for (r_{p}=0.13), (K=1.43), (n=0.0095), (alpha =1.15), (beta =0.99), (delta =0.2), (D=0.01), (Q=0.01), and variable correlation between additive and multiplicative noises: (a) (lambda =1), (b) (lambda =0.5) and (c) (lambda =0).Full size imageThe sustainability of the cell population depends on the strength of the two noises, like the correlation strength between them. For the zero strength multiplicative noise, the population has the mode at around the conditional MSSCD value (Fig. 11). Therefore, the population sustains in this case and tries to stabilize at the conditional MSSCD value. For (D=0.02), there is a bimodality, where the highest mode is at the zero cell density. For (D=0.05), we observe only one mode at (x=0). Therefore, with the increasing values of the multiplicative noise strengths (D), the chance of extinction increases for (lambda =0.5), (Q=0.01), and other constant model parameters for the seeding condition 1. Similar things happen for increasing Q values considering (D=0.01), (lambda =0.5), and other constant model parameters (Fig. 12).Figure 11Distribution of cell density for (r_{p}=0.13), (K=1.43), (n=0.0095), (alpha =1.15), (beta =0.99), (delta =0.2), (lambda =0.5), (Q=0.01), and variable strength of multiplicative noise: (a) (D=0.05), (b) (D=0.02) and (c) (D=0).Full size imageFigure 12Distribution of cell density for (r_{p}=0.13), (K=1.43), (n=0.0095), (alpha =1.15), (beta =0.99), (delta =0.2), (lambda =0.5), (D=0.01), and variable correlation between multiplicative noise: (a) (Q=0.05), (b) (Q=0.02) and (c) (Q=0).Full size image Remark 5 We have previously discussed the scenario for (alpha =delta) for deterministic case in Remark 4. It is important to understand the scenario under stochastic case too. For (alpha =delta) the proposed stochastic model 3 becomes (frac{dx(t)}{dt}=r_{p}x(t)^{(alpha +1)}left( 1-big (frac{x(t)}{K}big )^{beta }right) - nx(t)^{(alpha +1)}-x(t)^{(alpha +1)} epsilon (t)+ Gamma (t)). For this stochastic model (g_{1}(x)=-x^{alpha +1}) and (g_{2}(x)=1). We get, (A(x)=r_{p}x^{alpha +1}left( 1-Big (frac{x}{K}Big )^{beta } right) -nx^{(alpha +1)}+D(alpha +1)x^{(2alpha +1)}-lambda sqrt{DQ}(alpha +1)x^{alpha }) and (B(x)=Dx^{(2alpha +2)}-2lambda sqrt{DQ}x^{(alpha +1)}+Q). The extrema of the SPDF (big (x(t)=x^{*}big )) must satisfy the growth equation (r_{p}{x^{*}}^{alpha +1}-frac{r_{p}}{K^{beta }}(x^{*})^{alpha +beta +1}-n(x^{*})^{alpha +1}-D(alpha +1)(x^{*})^{2alpha +1}+lambda sqrt{D~Q}(alpha +1)(x^{*})^{alpha }=0). Therefore, for (alpha =delta) the conditional MSSCD is (x^{*}= K-Kfrac{nK^{(alpha +1)}+D(alpha +1)K^{(2alpha +1)}-lambda sqrt{DQ}(alpha +1)K^{alpha }}{beta r_{p}K^{(alpha +1)}+nK^{(alpha +1)}(alpha +1)+D(alpha +1)(2alpha +1)K^{(2alpha +1)}-alpha lambda sqrt{DQ}(alpha +1)K^{alpha }}) under the condition ((r_{p}-n)(alpha +1)(x^{*})^{alpha }-frac{r_{p}}{K^{beta }}(alpha +beta +1)(x^{*})^{(alpha + beta )}-(alpha +1)(2alpha +1)D(x^{*})^{2alpha }+lambda sqrt{DQ}(alpha +1)alpha (x^{*})^{(alpha -1)} More

  • in

    Whales from space dataset, an annotated satellite image dataset of whales for training machine learning models

    Very high-resolution (VHR) satellite imagery allows us to survey regularly remote and large areas of the ocean, difficult to access by boats or planes. The interest in using VHR satellite imagery for the study of great whales (including sperm whales and baleen whales) has grown in the past years1,2,3,4,5 since Abileah6 and Fretwell et al.7 showed its potential. This growing interest may be linked to the improvement in the spatial resolution of satellite imagery, which increased in 2014 from 46 cm to 31 cm. This upgrade enhanced the confidence in the detection of whales in satellite imagery, as more details could be seen, such as whale-defining features (e.g. flukes).Detecting whales in the imagery is either conducted manually1,4,5,7, or automatically2,3. A downside of the manual approach is that it is time-demanding, with manual counter often having to view hundred and sometimes thousands of square kilometres of open ocean. The development of automated approaches to detect whales by satellite would not only speed up this application, but also reduce the possibility of missing whales due to observer fatigue and standardize the procedure. Various automated approaches exist from pixel-based to artificial intelligence. Machine learning, an application of artificial intelligence, seems to be the most appropriate automated method to detect whales efficiently in satellite imagery2,3,8,9.In machine learning an algorithm learns how to identify features by repeatedly testing different search parameters against a training dataset10,11. Concerning whales, the algorithm needs to be trained to detect the wide variety of shapes and colour characterising whales. Shapes and colour will be influenced by the type of species, the environment (e.g. various degree of turbidity), the light conditions, and the behaviours (e.g. foraging, travelling, breaching), as different behaviours will result in different postures. The larger a training dataset is, the more accurate and transferable to other satellite images the algorithm will be. At the time of writing, such a dataset does not exist or is not publicly available.Creating a large enough dataset necessary to train algorithms to detect whales in VHR satellite imagery will require the various research groups analysing VHR satellite imagery to openly share examples of whales and non-whale objects in VHR satellite imagery, which could be facilitated by uploading such data on a central open source repository, similar to the GenBank12 for DNA code or OBIS-Seamap13 for marine wildlife observations. Ideally clipped out image chips of the whale objects would be shared as tiff files, which retains most of the characteristics of the original image. However, all VHR satellites are commercially owned, except for the Cartosat-3 owned by the government of India14, which means it is not possible to publicly share image chips as tiff file. Instead, image chips could be shared in a png or jepg format, which involve loosing some spectral information. If tiff files are required, georeferenced and labelled boxes encompassing the whale objects could also be shared, including information on the satellite imagery to allow anyone to ask the commercial providers for the exact imagery.Here we present a database of whale objects found in VHR satellite imagery. It represents four different species of whales (i.e. southern right whale, Eubalaena australis; grey whale, Eschrichtius robustus; humpback whale, Megaptera novaeangliae; fin whale, Balaenoptera physalus; Fig. 1), which were manually detected in images captured by different satellites (i.e., GeoEye-1, Quickbird-2, WorldView-2, WorldView-3). We created the database by (i) first detecting whale objects manually in satellite imagery, (ii) then we classified whale objects as either “definite”, “probable” or “possible” as in Cubaynes et al.1; and (iii) finally we created georeferenced and labelled points and boxes centered around each whale object, as well as providing image chips in a png format. With this database made publicly available, we aim to initiate the creation of a central database that can be built upon.Fig. 1Database of annotated whales detected in satellite imagery covering different species and areas. Humpback whales were detected in Maui Nui, US (a); grey whales in Laguna San Ignacio, Mexico (b); fin whales in the Pelagos Sanctuary, France, Monaco and Italy (c); southern right whales were observed in three areas, off the Peninsula Valdes, Argentina (d); off Witsand, South Africa (e); and off the Auckland Islands, New Zealand (f). The dot size represents the number of annotated whales per location. Whale silhouettes were sourced from philopic.com (the grey and humpback whales silhouettes are from Chris Luh).Full size image More

  • in

    Increased abundance of a common scavenger affects allocation of carrion but not efficiency of carcass removal in the Fukushima Exclusion Zone

    Lim, N., Kelt, D. A., Lim, K. K. & Bernard, H. Vertebrate scavengers control abundance of diarrheal-causing bacteria in tropical plantations. Zool. Stud. 59, 1–10 (2020).
    Google Scholar 
    Beasley, J. C., Olson, Z. H. & DeVault, T. L. Ecological role of vertebrate scavengers. In: Carrion Ecology, Evolution and their Applications. (eds Benbow, E.M., Tomberlin, J. & Tarone, A.) 107–127 (CRC Press, 2015).
    Ogada, D. L., Keesing, F. & Virani, M. Z. Dropping dead: Causes and consequences of vulture population declines worldwide. Ann. N. Y. Acad. Sci. 1249, 57–71 (2012).ADS 
    PubMed 
    Article 

    Google Scholar 
    Reid, W. V. et al. Ecosystems and Human Well-Being-Synthesis: A Report of the Millennium Ecosystem Assessment (Island Press, 2005).
    Google Scholar 
    Wilson, E. E. & Wolkovich, E. M. Scavenging: How carnivores and carrion structure communities. Trends Ecol. Evol. 26, 129–135 (2011).PubMed 
    Article 

    Google Scholar 
    Moleón, M., Sánchez-Zapata, J. A., Selva, N., Donázar, J. A. & Owen-Smith, N. Inter-specific interactions linking predation and scavenging in terrestrial vertebrate assemblages. Biol. Rev. 89, 1042–1054. https://doi.org/10.1111/brv.12097 (2014).Article 
    PubMed 

    Google Scholar 
    Fonseca, C. R. & Ganade, G. Species functional redundancy, random extinctions and the stability of ecosystems. J. Ecol. 89, 118–125 (2001).Article 

    Google Scholar 
    Mori, A. S., Furukawa, T. & Sasaki, T. Response diversity determines the resilience of ecosystems to environmental change. Biol. Rev. 88, 349–364. https://doi.org/10.1111/brv.12004 (2013).Article 
    PubMed 

    Google Scholar 
    Huijbers, C. M. et al. Limited functional redundancy in vertebrate scavenger guilds fails to compensate for the loss of raptors from urbanized sandy beaches. Divers. Distrib. 21, 55–63 (2015).Article 

    Google Scholar 
    Ceballos, G. et al. Accelerated modern human–induced species losses: Entering the sixth mass extinction. Sci. Adv. 1, e1400253 (2015).ADS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Buechley, E. R. & Şekercioğlu, Ç. H. The Avian scavenger crisis: Looming extinctions, trophic cascades, and loss of critical ecosystem functions. Biol. Cons. 198, 220–228 (2016).Article 

    Google Scholar 
    Hill, J. E., DeVault, T. L., Wang, G. & Belant, J. L. Anthropogenic mortality in mammals increases with the human footprint. Front. Ecol. Environ. 18, 13–18. https://doi.org/10.1002/fee.2127 (2019).Article 

    Google Scholar 
    Sebastián-González, E. et al. Scavenging in the Anthropocene: Human impact drives vertebrate scavenger species richness at a global scale. Glob. Change Biol. 25, 3005–3017 (2019).ADS 
    Article 

    Google Scholar 
    Sebastián-González, E. et al. Network structure of vertebrate scavenger assemblages at the global scale: Drivers and ecosystem functioning implications. Ecography 43, 1–13. https://doi.org/10.1111/ecog.05083 (2020).Article 

    Google Scholar 
    Marneweck, C. J., Katzner, T. E. & Jachowski, D. S. Predicted climate-induced reductions in scavenging in eastern North America. Glob. Change Biol. 27, 3383–3394. https://doi.org/10.1111/gcb.15653 (2021).Article 

    Google Scholar 
    Mokany, K., Ash, J. & Roxburgh, S. Functional identity is more important than diversity in influencing ecosystem processes in a temperate native grassland. J. Ecol. 96, 884–893. https://doi.org/10.1111/j.1365-2745.2008.01395.x (2008).Article 

    Google Scholar 
    Gagic, V. et al. Functional identity and diversity of animals predict ecosystem functioning better than species-based indices. Proc. R. Soc. B Biol. Sci. 282, 20142620 (2015).Article 

    Google Scholar 
    Mateo-Tomás, P., Olea, P. P., Selva, N. & Sánchez-Zapata, J. A. Species and individual replacements contribute more than nestedness to shape vertebrate scavenger metacommunities. Ecography 42, 365–375 (2019).Article 

    Google Scholar 
    Sebastián-González, E. et al. Functional traits driving species role in the structure of terrestrial vertebrate scavenger networks. Ecology https://doi.org/10.1002/ecy.3519 (2021).Article 
    PubMed 

    Google Scholar 
    DeVault, T. L., Rhodes, O. E. Jr. & Shivik, J. A. Scavenging by vertebrates: Behavioral, ecological, and evolutionary perspectives on an important energy transfer pathway in terrestrial ecosystems. Oikos 102, 225–234 (2003).Article 

    Google Scholar 
    Allen, M. L., Elbroch, L. M., Wilmers, C. C. & Wittmer, H. U. The comparative effects of large carnivores on the acquisition of carrion by scavengers. Am. Nat. 185, 822–833 (2015).PubMed 
    Article 

    Google Scholar 
    Moleón, M., Sánchez-Zapata, J. A., Sebastián-González, E. & Owen-Smith, N. Carcass size shapes the structure and functioning of an African scavenging assemblage. Oikos 124, 1391–1403 (2015).Article 

    Google Scholar 
    Gutiérrez-Cánovas, C. et al. Large home range scavengers support higher rates of carcass removal. Funct. Ecol. 34, 1921–1932 (2020).Article 

    Google Scholar 
    Walker, M. A. et al. Factors influencing scavenger guilds and scavenging efficiency in Southwestern Montana. Sci. Rep. https://doi.org/10.1038/s41598-021-83426-3 (2021).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Winfree, R., Fox, J., Williams, N. M., Reilly, J. R. & Cariveau, D. P. Abundance of common species, not species richness, drives delivery of a real-world ecosystem service. Ecol. Lett. 18, 626–635. https://doi.org/10.1111/ele.12424 (2015).Article 
    PubMed 

    Google Scholar 
    Mateo-Tomás, P., Olea, P. P., Moleón, M., Selva, N. & Sánchez-Zapata, J. A. Both rare and common species support ecosystem services in scavenger communities. Glob. Ecol. Biogeogr. 26, 1459–1470. https://doi.org/10.1111/geb.12673 (2017).Article 

    Google Scholar 
    Butler, J. R. A. & du Toit, J. T. Diet of free-ranging domestic dogs (Canis familiaris) in rural Zimbabwe: Implications for wild scavengers on the periphery of wildlife reserves. Anim. Conserv. 5, 29–37. https://doi.org/10.1017/s136794300200104x (2002).Article 

    Google Scholar 
    DeVault, T. L., Olson, Z. H., Beasley, J. C. & Rhodes, O. E. Jr. Mesopredators dominate competition for carrion in an agricultural landscape. Basic Appl. Ecol. 12, 268–274 (2011).Article 

    Google Scholar 
    Ogada, D. L., Torchin, M. E., Kinnaird, M. F. & Ezenwa, V. O. Effects of vulture declines on facultative scavengers and potential implications for mammalian disease transmission. Conserv. Biol. 26, 453–460. https://doi.org/10.1111/j.1523-1739.2012.01827.x (2012).CAS 
    Article 
    PubMed 

    Google Scholar 
    Morales-Reyes, Z. et al. Scavenging efficiency and red fox abundance in Mediterranean mountains with and without vultures. Acta Oecol. 79, 81–88. https://doi.org/10.1016/j.actao.2016.12.012 (2017).ADS 
    Article 

    Google Scholar 
    Inagaki, A. et al. Vertebrate scavenger guild composition and utilization of carrion in an East Asian temperate forest. Ecol. Evol. 10, 1223–1232 (2020).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Blazquez, M., Sanchez-Zapata, J. A., Botella, F., Carrete, M. & Eguía, S. Spatio-temporal segregation of facultative avian scavengers at ungulate carcasses. Acta Oecol. 35, 645–650 (2009).ADS 
    Article 

    Google Scholar 
    Inger, R., Cox, D. T. C., Per, E., Norton, B. A. & Gaston, K. J. Ecological role of vertebrate scavengers in urban ecosystems in the UK. Ecol. Evol. 6, 7015–7023. https://doi.org/10.1002/ece3.2414 (2016).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Hill, J. E., DeVault, T. L., Beasley, J. C., Rhodes, O. E. Jr. & Belant, J. L. Effects of vulture exclusion on carrion consumption by facultative scavengers. Ecol. Evol. 8, 2518–2526. https://doi.org/10.1002/ece3.3840 (2018).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Olson, Z., Beasley, J., DeVault, T. L. & Rhodes, O. E. Jr. Scavenger community response to the removal of a dominant scavenger. Oikos 121, 77–84 (2012).Article 

    Google Scholar 
    Pardo-Barquín, E., Mateo-Tomás, P. & Olea, P. P. Habitat characteristics from local to landscape scales combine to shape vertebrate scavenging communities. Basic Appl. Ecol. 34, 126–139. https://doi.org/10.1016/j.baae.2018.08.005 (2019).Article 

    Google Scholar 
    Turner, K. L., Conner, L. M. & Beasley, J. C. Effect of mammalian mesopredator exclusion on vertebrate scavenging communities. Sci. Rep. 10, 1–9 (2020).Article 
    CAS 

    Google Scholar 
    Ohashi, H. et al. Differences in the activity pattern of the wild boar Sus scrofa related to human disturbance. Eur. J. Wildl. Res. 59, 167–177. https://doi.org/10.1007/s10344-012-0661-z (2013).Article 

    Google Scholar 
    Saito, M. & Koike, F. Distribution of wild mammal assemblages along an urban–rural–forest landscape gradient in warm-temperate East Asia. PLoS ONE 8, e65464. https://doi.org/10.1371/journal.pone.0065464 (2013).ADS 
    CAS 
    Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Gaynor, K. M., Hojnowski, C. E., Carter, N. H. & Brashares, J. S. The influence of human disturbance on wildlife nocturnality. Science 360, 1232–1235. https://doi.org/10.1126/science.aar7121 (2018).ADS 
    CAS 
    Article 
    PubMed 

    Google Scholar 
    Tsunoda, M. et al. Human disturbance affects latrine-use patterns of raccoon dogs. J. Wildl. Manag. 83, 728–736. https://doi.org/10.1002/jwmg.21610 (2019).Article 

    Google Scholar 
    Watabe, R. & Saito, M. U. Effects of vehicle-passing frequency on forest roads on the activity patterns of carnivores. Landsc. Ecol. Eng. 17, 225–231. https://doi.org/10.1007/s11355-020-00434-7 (2021).Article 

    Google Scholar 
    Luna, Á., Romero-Vidal, P. & Arrondo, E. Predation and scavenging in the city: A review of spatio-temporal trends in research. Diversity 13, 46. https://doi.org/10.3390/d13020046 (2021).Article 

    Google Scholar 
    Huijbers, C. M., Schlacher, T. A., Schoeman, D. S., Weston, M. A. & Connolly, R. M. Urbanisation alters processing of marine carrion on sandy beaches. Landsc. Urban Plan. 119, 1–8 (2013).Article 

    Google Scholar 
    Fukushima Prefectural Government. Transition of evacuation designated zones. https://www.pref.fukushima.lg.jp/site/portal-english/en03-08.html. (2019). Accessed 20 Apr 2022.Steinhauser, G., Brandl, A. & Johnson, T. E. Comparison of the Chernobyl and Fukushima nuclear accidents: A review of the environmental impacts. Sci. Total Environ. 470, 800–817 (2014).ADS 
    PubMed 
    Article 
    CAS 

    Google Scholar 
    Center for International Earth Science Information Network (CIESIN)—Columbia University. (NASA Socioeconomic Data and Applications Center (SEDAC), Palisades, NY, 2018).Lyons, P. C., Okuda, K., Hamilton, M. J., Hinton, T. G. & Beasley, J. C. Rewilding of Fukushima’s human evacuation zone in the presence of radioactive stressors. Front. Ecol. Environ. 18, 127–134 (2020).Article 

    Google Scholar 
    Deryabina, T. G. et al. Long-term census data reveal abundant wildlife populations at Chernobyl. Curr. Biol. 25, R824–R826. https://doi.org/10.1016/j.cub.2015.08.017 (2015).CAS 
    Article 
    PubMed 

    Google Scholar 
    Webster, S. C. et al. Where the wild things are: Influence of radiation on the distribution of four mammalian species within the Chernobyl Exclusion Zone. Front. Ecol. Environ. 14, 185–190. https://doi.org/10.1002/fee.1227 (2016).Article 

    Google Scholar 
    Schlichting, P. E., Love, C. N., Webster, S. C. & Beasley, J. C. Efficiency and composition of vertebrate scavengers at the land–water interface in the Chernobyl Exclusion Zone. Food Webs 18, e00107. https://doi.org/10.1016/j.fooweb.2018.e00107 (2019).Article 

    Google Scholar 
    Newsome, T. M. et al. Monitoring the dead as an ecosystem indicator. Ecol. Evol. 11, 5844–5856. https://doi.org/10.1002/ece3.7542 (2021).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Turner, K. L., Abernethy, E. F., Mike Conner, L., Rhodes, O. E. Jr. & Beasley, J. C. Abiotic and biotic factors modulate carrion fate and vertebrate scavenging communities. Ecology 98, 2413–2424 (2017).PubMed 
    Article 

    Google Scholar 
    Ruzicka, R. E. & Conover, M. R. Does weather or site characteristics influence the ability of scavengers to locate food?. Ethology 118, 187–196 (2012).Article 

    Google Scholar 
    Paula, J. J. S. et al. Camera-trapping as a methodology to assess the persistence of wildlife carcasses resulting from collisions with human-made structures. Wildl. Res. 41, 717–725. https://doi.org/10.1071/WR14063 (2015).Article 

    Google Scholar 
    Selva, N., Jędrzejewska, B., Jędrzejewski, W. & Wajrak, A. Factors affecting carcass use by a guild of scavengers in European temperate woodland. Can. J. Zool. 83, 1590–1601 (2005).Article 

    Google Scholar 
    Nakama, S., Yoshimura, K., Fujiwara, K., Ishikawa, H. & Iijima, K. Temporal decrease in air dose rate in the sub-urban area affected by the Fukushima Dai-ichi Nuclear Power Plant accident during four years after decontamination works. J. Environ. Radioact. 208–209, 106013. https://doi.org/10.1016/j.jenvrad.2019.106013 (2019).CAS 
    Article 
    PubMed 

    Google Scholar 
    Ministry of the Environment of Japan. Off-Site Environmental Remediation in Affected Areas in Japan. http://josen.env.go.jp/en/decontamination/ (2020). Accessed 20 Apr 2022.Japan Meteorological Agency. Climate in Namie in 2018: Monthly Overview Data. http://www.data.jma.go.jp/obd/stats/etrn/view/monthly_a1.php?prec_no=36&block_no=0295&year=2018&month=7&day=&view=p1 (2018). Accessed 1 Apr 2019.De Vault, T. L., Brisbin, J., Lehr, I., Rhodes, J. & Olin, E. Factors influencing the acquisition of rodent carrion by vertebrate scavengers and decomposers. Can. J. Zool. 82, 502–509 (2004).Article 

    Google Scholar 
    Kane, A., Healy, K., Guillerme, T., Ruxton, G. D. & Jackson, A. L. A recipe for scavenging in vertebrates—The natural history of a behaviour. Ecography 40, 11. https://doi.org/10.1111/ecog.02817 (2017).Article 

    Google Scholar 
    Natusch, D. J. D., Lyons, J. A. & Shine, R. How do predators and scavengers locate resource hotspots within a tropical forest?. Aust. Ecol. 42, 742–749. https://doi.org/10.1111/aec.12492 (2017).Article 

    Google Scholar 
    Japan Aerospace Exploration Agency. High-resolution land use land cover map of Japan (ver.16.09). https://www.eorc.jaxa.jp/ALOS/en/lulc/lulc_index.htm (2011). Accessed 1 Apr 2019.Newkirk, E. S. CPW Photo Warehouse. http://cpw.state.co.us/learn/Pages/ResearchMammalsSoftware.aspx (2016). Accessed 1 Apr 2019.Therneau, T. M. A Package for Survival Analysis in R. R package version 3.3-1 (2022).Bates, D., Mächler, M., Bolker, B. & Walker, S. Fitting linear mixed-effects models using lme4. J. Stat. Softw. 67, 1–48. https://doi.org/10.18637/jss.v067.i01 (2015).Article 

    Google Scholar 
    Anderson, D. et al. Introgression dynamics from invasive pigs into wild boar following the March 2011 natural and anthropogenic disasters at Fukushima. Proc. R. Soc. B Biol. Sci. 288, 20210874. https://doi.org/10.1098/rspb.2021.0874 (2021).CAS 
    Article 

    Google Scholar 
    Ishiniwa, H., Onuma, M. & Tamaoki, M. Behavior of Radionuclides in the Environment III 463–472 (Springer, 2022).Book 

    Google Scholar 
    Nemoto, Y. et al. Effects of 137Cs contamination after the TEPCO Fukushima Dai-ichi Nuclear Power Station accident on food and habitat of wild boar in Fukushima Prefecture. J. Environ. Radioact. 225, 106342. https://doi.org/10.1016/j.jenvrad.2020.106342 (2020).CAS 
    Article 
    PubMed 

    Google Scholar 
    Olson, Z. H., Beasley, J. C. & Rhodes, O. E. Jr. Carcass type affects local scavenger guilds more than habitat connectivity. PLoS ONE 11, e0147798 (2016).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    DeVault, T. L., Seamans, T. W., Linnell, K. E., Sparks, D. W. & Beasley, J. C. Scavenger removal of bird carcasses at simulated wind turbines: Does carcass type matter?. Ecosphere. https://doi.org/10.1002/ecs2.1994 (2017).Article 

    Google Scholar 
    Sugiura, S., Tanaka, R., Taki, H. & Kanzaki, N. Differential responses of scavenging arthropods and vertebrates to forest loss maintain ecosystem function in a heterogeneous landscape. Biol. Cons. 159, 206–213 (2013).Article 

    Google Scholar 
    Enari, H. & Enari, H. S. Not avian but mammalian scavengers efficiently consume carcasses under heavy snowfall conditions: A case from northern Japan. Mamm. Biol. 101, 419–428. https://doi.org/10.1007/s42991-020-00097-9 (2021).Article 

    Google Scholar 
    Selva, N., Jedrzejewska, B., Jedrzejewski, W. & Wajrak, A. Scavenging on European bison carcasses in Bialowieza primeval forest (eastern Poland). Ecoscience 10, 303–311 (2003).Article 

    Google Scholar 
    Jojola-Elverum, S. M., Shivik, J. A. & Clark, L. Importance of bacterial decomposition and carrion substrate to foraging brown treesnakes. J. Chem. Ecol. 27, 1315–1331. https://doi.org/10.1023/a:1010357024140 (2001).CAS 
    Article 
    PubMed 

    Google Scholar 
    Abernethy, E. F., Turner, K. L., Beasley, J. C. & Rhodes, O. E. Jr. Scavenging along an ecological interface: Utilization of amphibian and reptile carcasses around isolated wetlands. Ecosphere 8, e01989. https://doi.org/10.1002/ecs2.1989 (2017).Article 

    Google Scholar 
    Sugiura, S. & Hayashi, M. Functional compensation by insular scavengers: The relative contributions of vertebrates and invertebrates vary among islands. Ecography 41, 1173–1183 (2018).Article 

    Google Scholar 
    Matsuo, R. & Ochiai, K. Dietary overlap among two introduced and one native sympatric carnivore species, the raccoon, the masked palm civet, and the raccoon dog, in Chiba Prefecture, Japan. Mammal Study 34, 187–194 (2009).Article 

    Google Scholar 
    Drygala, F. & Zoller, H. Diet composition of the invasive raccoon dog (Nyctereutes procyonoides) and the native red fox (Vulpes vulpes) in north-east Germany. Hystrix Italian J. Mammal. 24, 190–194 (2014).
    Google Scholar 
    Elmeros, M. et al. The diet of feral raccoon dog (Nyctereutes procyonoides) and native badger (Meles meles) and red fox (Vulpes vulpes) in Denmark. Mammal Res. 63, 405–413. https://doi.org/10.1007/s13364-018-0372-2 (2018).Article 

    Google Scholar 
    Sekizawa, R., Ichii, K. & Kondo, M. Satellite-based detection of evacuation-induced land cover changes following the Fukushima Daiichi nuclear disaster. Remote Sensing Lett. 6, 824–833 (2015).Article 

    Google Scholar 
    Ishihara, M. & Tadono, T. Land cover changes induced by the great east Japan earthquake in 2011. Sci. Rep. 7, 45769–45769. https://doi.org/10.1038/srep45769 (2017).ADS 
    Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Focardi, S., Materassi, M., Innocenti, G. & Berzi, D. Kleptoparasitism and scavenging can stabilize ecosystem dynamics. Am. Nat. 190, 398–409 (2017).PubMed 
    Article 

    Google Scholar 
    Osugi, S., Trentin, B. E. & Koike, S. Impact of wild boars on the feeding behavior of smaller frugivorous mammals. Mamm. Biol. 97, 22–27 (2019).Article 

    Google Scholar 
    Duľa, M. & Krofel, M. A cat in paradise: Hunting and feeding behaviour of Eurasian lynx among abundant naive prey. Mamm. Biol. 100, 685–690. https://doi.org/10.1007/s42991-020-00070-6 (2020).Article 

    Google Scholar 
    Smith, J. B., Laatsch, L. J. & Beasley, J. C. Spatial complexity of carcass location influences vertebrate scavenger efficiency and species composition. Sci. Rep. 7, 10250. https://doi.org/10.1038/s41598-017-10046-1 (2017).ADS 
    CAS 
    Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Moleón, M. et al. Carrion availability in space and time. In Carrion Ecology and Management (eds Olea, P.P., Mateo-Tomás, P. & Sánchez-Zapata, J.A.) 23–44 (Springer International Publishing, 2019).
    DeVault, T. L. & Rhodes, O. E. Jr. Identification of vertebrate scavengers of small mammal carcasses in a forested landscape. Acta Theriol. 47, 185–192 (2002).Article 

    Google Scholar 
    Bumann, G. B. & Stauffer, D. F. Scavenging of ruffed grouse in the Appalachians: Influences and implications. Wildl. Soc. Bull. 1973–2006(30), 853–860 (2002).
    Google Scholar 
    Young, A., Stillman, R., Smith, M. J. & Korstjens, A. H. An experimental study of vertebrate scavenging behavior in a Northwest European woodland context. J. Forensic Sci. 59, 1333–1342. https://doi.org/10.1111/1556-4029.12468 (2014).Article 
    PubMed 

    Google Scholar 
    Abernethy, E. F. et al. Carcasses of invasive species are predominantly utilized by invasive scavengers in an island ecosystem. Ecosphere 7 (2016).DeVault, T. L. & Krochmal, A. R. Scavenging by snakes: An examination of the literature. Herpetologica 58, 429–436 (2002).Article 

    Google Scholar 
    Shivik, J. A. & Clark, L. Ontogenetic shifts in carrion attractiveness to brown tree snakes (Boiga irregularis). J. Herpetol. 33, 334–336. https://doi.org/10.2307/1565737 (1999).Article 

    Google Scholar 
    Campobasso, C. P., Di Vella, G. & Introna, F. Factors affecting decomposition and Diptera colonization. Forensic Sci. Int. 120, 18–27 (2001).CAS 
    PubMed 
    Article 

    Google Scholar  More

  • in

    Reply to: Assessing the efficiency of Verily’s automated process for production and release of male Wolbachia-infected mosquitoes

    Crawford, J. E. et al. Efficient production of male Wolbachia-infected Aedes aegypti mosquitoes enables large-scale suppression of wild populations. Nat. Biotechnol. 38, 482–492 (2020).CAS 
    Article 

    Google Scholar 
    Xi, Z., Khoo, C. C. H. & Dobson, S. L. Wolbachia establishment and invasion in an Aedes aegypti laboratory population. Science 310, 326–328 (2005).CAS 
    Article 

    Google Scholar 
    Phuc, H. K. et al. Late-acting dominant lethal genetic systems and mosquito control. BMC Biol. 5, 11 (2007).Article 

    Google Scholar 
    Kandul, N. P. et al. Transforming insect population control with precision guided sterile males with demonstration in flies. Nat. Commun. 10, 84 (2019).CAS 
    Article 

    Google Scholar 
    Kyrou, K. et al. A CRISPR–Cas9 gene drive targeting doublesex causes complete population suppression in caged Anopheles gambiae mosquitoes. Nat. Biotechnol. 36, 1062–1066 (2018).CAS 
    Article 

    Google Scholar 
    Kittayapong, P. et al. Combined sterile insect technique and incompatible insect technique: the first proof-of-concept to suppress Aedes aegypti vector populations in semi-rural settings in Thailand. PLoS Negl. Trop. Dis. 13, e0007771 (2019).Article 

    Google Scholar 
    Zheng, X. et al. Incompatible and sterile insect techniques combined eliminate mosquitoes. Nature 572, 56–61 (2019).CAS 
    Article 

    Google Scholar 
    Ryan, P. A. et al. Establishment of wMel Wolbachia in Aedes aegypti mosquitoes and reduction of local dengue transmission in Cairns and surrounding locations in northern Queensland, Australia. Gates Open Res. 3, 1547 (2019).Article 

    Google Scholar 
    Indriani, C. et al. Reduced dengue incidence following deployments of Wolbachia-infected Aedes aegypti in Yogyakarta, Indonesia: a quasi-experimental trial using controlled interrupted time series analysis. Gates Open Res. 4, 50 (2020).Velez, I. D. et al. The impact of city-wide deployment of Wolbachia-carrying mosquitoes on arboviral disease incidence in Medellín and Bello, Colombia: study protocol for an interrupted time-series analysis and a test-negative design study. F1000Res. 8, 1327 (2020).Article 

    Google Scholar 
    Durovni, B. et al. The impact of large-scale deployment of Wolbachia mosquitoes on dengue and other Aedes-borne diseases in Rio de Janeiro and Niterói, Brazil: study protocol for a controlled interrupted time series analysis using routine disease surveillance data. F1000Res. 8, 1328 (2020).Article 

    Google Scholar 
    O’Connor, L. et al. Open release of male mosquitoes infected with a Wolbachia biopesticide: field performance and infection containment. PLoS Negl. Trop. Dis. 6, e1797 (2012).Article 

    Google Scholar 
    Nazni, W. A. et al. Establishment of Wolbachia strain wAlbB in Malaysian populations of Aedes aegypti for dengue control. Curr. Biol. 29, 4241–4248 (2019).CAS 
    Article 

    Google Scholar 
    Klassen, W. & Curtis, C. F. In: Sterile Insect Technique (eds Dyck, V. A., Hendrichs, J. & Robinson, A. S.) 3–36 (Springer-Verlag, 2005).Fried, M. Determination of sterile-insect competitiveness. J. Econ. Entomol. 64, 869–872 (1971).Article 

    Google Scholar 
    Bouyer, J. et al. Field performance of sterile male mosquitoes released from an uncrewed aerial vehicle. Sci. Robot. 5, eaba6251 (2020).Article 

    Google Scholar 
    Krafsur, E. S., Whitten, C. J. & Novy, J. E. Screwworm eradication in North and Central America. Parasitol. Today 3, 131–137 (1987).CAS 
    Article 

    Google Scholar 
    Hendrichs, J., Ortiz, G., Liedo, P. & Schwarz, A. Six years of successful medfly program in Mexico and Guatemala. In: Fruit Flies of Economic Importance (ed Cavalloro, R.) 353–365 (A. A. Balkema, 1983).Helinski, M. E. H., Parker, A. G. & Knols, B. G. J. Radiation-induced sterility for pupal and adult stages of the malaria mosquito Anopheles arabiensis. Malar. J. 5, 41 (2006).Article 

    Google Scholar 
    Helinski, M. E. H., Parker, A. G. & Knols, B. G. J. Radiation biology of mosquitoes. Malar. J. 8 Suppl 2, S6 (2009).Benedict, M. Q. & Robinson, A. S. The first releases of transgenic mosquitoes: an argument for the sterile insect technique. Trends Parasitol. 19, 349–355 (2003).Article 

    Google Scholar 
    Culbert, N. J. et al. Longevity of mass-reared, irradiated and packed male Anopheles arabiensis and Aedes aegypti under simulated environmental field conditions. Parasit. Vectors 11, 603 (2018).CAS 
    Article 

    Google Scholar 
    Culbert, N. J. et al. A rapid quality control test to foster the development of genetic control in mosquitoes. Sci. Rep. 8, 16179 (2018).Article 

    Google Scholar 
    Bentley, D. R. et al. Accurate whole human genome sequencing using reversible terminator chemistry. Nature 456, 53–59 (2008).CAS 
    Article 

    Google Scholar 
    Carlson, R. The pace and proliferation of biological technologies. Biosecur. Bioterror. 1, 203–214 (2003).Article 

    Google Scholar 
    The Wolbachia Project–Singapore Consortium & Ching, N. L. Wolbachia-mediated sterility suppresses Aedes aegypti populations in the urban tropics. Preprint at https://www.medrxiv.org/content/10.1101/2021.06.16.21257922v1 (2021).Soh, S. et al. Economic impact of dengue in Singapore from 2010 to 2020 and the cost-effectiveness of Wolbachia interventions. PLoS Global Public Health https://doi.org/10.1371/journal.pgph.0000024 (2021). More

  • in

    Microbiomes of microscopic marine invertebrates do not reveal signatures of phylosymbiosis

    Gilbert, S. F., Sapp, J. & Tauber, A. I. A symbiotic view of life: we have never been individuals. Q. Rev. Biol. 87, 325–341 (2012).PubMed 
    Article 

    Google Scholar 
    Bass, D., Stentiford, G. D., Wang, H.-C., Koskella, B. & Tyler, C. R. The pathobiome in animal and plant diseases. Trends Ecol. Evol. 34, 996–1008 (2019).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Husnik, F. & Keeling, P. J. The fate of obligate endosymbionts: reduction, integration, or extinction. Curr. Opin. Genet. Dev. 58-59, 1–8 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Berg, G. et al. Microbiome definition re-visited: old concepts and new challenges. Microbiome 8, 103 (2020).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Kwong, W. K. & Moran, N. A. Gut microbial communities of social bees. Nat. Rev. Microbiol. 14, 374–384 (2016).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Hammer, T. J., Janzen, D. H., Hallwachs, W., Jaffe, S. P. & Fierer, N. Caterpillars lack a resident gut microbiome. Proc. Nat Acad. Sci. USA 114, 9641–9646 (2017).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Holt, C. C., van der Giezen, M., Daniels, C. L., Stentiford, G. D. & Bass, D. Spatial and temporal axes impact ecology of the gut microbiome in juvenile European lobster (Homarus gammarus). ISME J. 14, 531–543 (2020).PubMed 
    Article 

    Google Scholar 
    Pollock, F. J. et al. Coral-associated bacteria demonstrate phylosymbiosis and cophylogeny. Nat. Commun. 9, 4921 (2018).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Thomas, T. et al. Diversity, structure and convergent evolution of the global sponge microbiome. Nat. Commun. 7, 11870 (2016).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Engelberts, J. P. et al. Characterization of a sponge microbiome using an integrative genome-centric approach. ISME J. 14, 1100–1110 (2020).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Ley, R. E. et al. Evolution of mammals and their gut microbes. Science 320, 1647–1651 (2008).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Mallot, E. K. & Amato, K. R. Host specificity of the gut microbiome. Nat. Rev. Microbiol. 19, 639–653 (2021).Article 
    CAS 

    Google Scholar 
    Colston, T. J. & Jackson, C. R. Microbiome evolution along divergent branches of the vertebrate tree of life: what is known and unknown. Mol. Ecol. 25, 3776–3800 (2016).PubMed 
    Article 

    Google Scholar 
    Levin, D. et al. Diversity and functional landscapes in the microbiota of animals in the wild. Science 372, eabb5352 (2021).CAS 
    PubMed 
    Article 

    Google Scholar 
    Nishida, A. H. & Ochman, H. Rates of gut microbiome divergence in mammals. Mol. Ecol. 27, 1884–1897 (2013).Article 

    Google Scholar 
    Brooks, A. W., Kohl, K. D., Brucker, R. M., van Opstal, E. J. & Bordenstein, S. R. Phylosymbiosis: relationships and functional effects of microbial communities across host evolutionary history. PLoS Biol. 14, e2000225 (2016).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Mazel, F. et al. Is host filtering the main driver of phylosymbiosis across the tree of life? mSystems 3, https://doi.org/10.1128/mSystems.00097-18 (2018).Lutz, H. L. et al. Ecology and host identity outweigh evolutionary history in shaping the bat microbiome. mBio 4, 6 (2019).
    Google Scholar 
    Grond, K. et al. No evidence for phylosymbiosis in Western chipmunk species. FEMS Microbiol. Ecol. 96, fiz182 (2020).CAS 
    PubMed 
    Article 

    Google Scholar 
    Song, S. J. et al. Comparative analyses of vertebrate gut microbiomes reveal convergence between birds and bats. mBio 11, 1 (2020).Article 

    Google Scholar 
    Trevelline, B. K., Sosa, J., Hartup, B. K. & Kohl, K. D. A bird’s-eye view of phylosymbiosis: weak signatures of phylosymbiosis among all 15 species of cranes. Proc. R. Soc. B 287, 20192988 (2020).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Muegge, B. D. et al. Diet drives convergence in gut microbiome functions across mammalian phylogeny and within humans. Science 332, 970–974 (2011).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Youngblut, N. D. et al. Host diet and evolutionary history explain different aspects of gut microbiome diversity among vertebrate clades. Nat. Commun. 10, 2200 (2019).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Amato, K. R. et al. Evolutionary trends in host physiology outweigh dietary niche in structuring primate gut microbiomes. ISME J. 13, 576–587 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Moeller, A. H. et al. Social behavior shapes the chimpanzee pan-microbiome. Sci. Adv. 2, e1500997 (2016).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Eckert, E. M., Anicic, N. & Fontaneto, D. Freshwater zooplankton microbiome composition is highly flexible and strongly influenced by the environment. Mol. Ecol. 30, 1545–1558 (2021).PubMed 
    Article 

    Google Scholar 
    Yatsunenko, T. et al. Human gut microbiome viewed across age and geography. Nature 486, 222–228 (2012).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Bik, H. M. Microbial metazoa are microbes too. mSystems 4, e00109–e00119 (2019).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Schuelke, T., Pereira, T. J., Hardy, S. M. & Bik, H. M. Nematode-associated microbial taxa do not correlate with host phylogeny, geographic region or feeding morphology in marine sediment habitats. Mol. Ecol. 27, 1930–1951 (2018).PubMed 
    Article 

    Google Scholar 
    Guidetti, R. et al. Further insights in the Tardigrada microbiome: phylogenetic position and prevalence of infection of four new Alphaproteobacteria putative endosymbionts. Zool. J. Linn. Soc. 188, 925–937 (2020).Article 

    Google Scholar 
    Giere, O. Meiobenthology (Springer-Verlag, 2009).Laumer, C. E. et al. Revisiting metazoan phylogeny with genomic sampling of all phyla. Proc. R. Soc. B 286, 20190831 (2019).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Hammer, T. J., Sanders, J. G. & Fierer, N. Not all animals need a microbiome. FEMS Microbiol. Lett. 366, fnz117 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Alejandre-Colomo, C. et al. Cultivable Winogradskyella species are genomically distinct from the sympatric abundant candidate species. ISME Commun. 1, 51 (2021).Article 

    Google Scholar 
    Husnik, F. et al. Bacterial and archaeal symbioses with protists. Curr. Biol. 31, R862–R877 (2021).CAS 
    PubMed 
    Article 

    Google Scholar 
    Salje, J. Cells within cells: Rickettsiales and the obligate intracellular bacterial lifestyle. Nat. Rev. Microbiol. 19, 375–390 (2021).CAS 
    PubMed 
    Article 

    Google Scholar 
    Neave, M. J., Apprill, A., Ferrier-Pagès, C. & Voolstra, C. R. Diversity and function of prevalent symbiotic marine bacteria in the genus Endozoicomonas. Appl. Microbiol. Biotechnol. 100, 8315–8324 (2016).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Weiland-Bräuer, N. et al. Composition of bacterial communities associated with Aurelia aurita changes with compartment, life stage, and population. Appl. Environ. Microbiol. 81, 6038–6052 (2015).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Bik, E. M. et al. Marine mammals harbor unique microbiotas shaped by and yet distinct from the sea. Nat. Commun. 7, 10516 (2016).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Burns, A. R. et al. Contribution of neutral processes to the assembly of gut microbial communities in the zebrafish over host development. ISME J. 10, 655–664 (2016).CAS 
    PubMed 
    Article 

    Google Scholar 
    McFall-Ngai, M. Adaptive immunity: care for the community. Nature 445, 153 (2007).CAS 
    PubMed 
    Article 

    Google Scholar 
    Ruehland, C. & Dubilier, N. Gamma- and epsilonproteobacterial ectosymbionts of a shallow-water marine worm are related to deep-sea hydrothermal vent ectosymbionts. Environ. Microbiol. 12, 2312–2326 (2010).CAS 
    PubMed 

    Google Scholar 
    Gruber-Vodicka, H. R. et al. Two intracellular and cell-type specific bacterial symbionts in the placozoan Trichoplax H2. Nat. Microbiol. 4, 1465–1474 (2019).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Schockaert, E. R. in Methods for the Examination of Organismal Diversity in Soils and Sediments (ed. Hall, G. S.) 211–225 (CABI, 1996).Higgins, R. P. in Introduction to the Study of Meiofauna (eds. Higgins, R. P. and Thiel, H.) 328–331 (SIP, 1988).Schram, M. D. & Davison, P. G. Irwin Loops—a history and method of constructing homemade loops. Trans. Kans. Acad. Sci. 115, 35–40 (1903).Article 

    Google Scholar 
    Medlin, L., Elwood, H. J., Stickel, S. & Sogin, M. L. The characterization of enzymatically amplified eukaryotic 16S-like rRNA-coding regions. Gene 71, 491–499 (1988).CAS 
    PubMed 
    Article 

    Google Scholar 
    Bower, S. M. et al. Preferential PCR amplification of parasitic protistan small subunit rDNA from metazoan tissues. J. Eukaryot. Microbiol. 51, 325–332 (2004).CAS 
    PubMed 
    Article 

    Google Scholar 
    Comeau, A. M., Li, W. K. W., Tremblay, J.-E., Carmack, E. C. & Lovejoy, C. Arctic ocean microbial community structure before and after the 2007 record sea ice minimum. PLoS ONE 6, e27492 (2011).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Zhang, R.-Y. et al. Design of targeted primers based on 16S rRNA sequences in meta-transcriptomic datasets and identification of a novel taxonomic group in the Asgard archaea. BMC Microbiol. 20, 25 (2020).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Lane, D. J. in Nucleic Acid Techniques in Bacterial Systematics (eds Stackebrandt, E. & Goodfellow, M) 115–175 (Wiley, 1991).Parada, A. E., Needham, D. M. & Fuhrman, J. A. Every base matters: assessing small subunit rRNA primers for marine microbiomes with mock communities, time series and global field samples. Environ. Microbiol. 18, 1403–1414 (2016).CAS 
    PubMed 
    Article 

    Google Scholar 
    Marcel, M. Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet J. 17, 10 (2011).
    Google Scholar 
    R Core Team. R: A Language and Environment for Statistical Computing (R Foundation for Statistical Computing, 2020).Callahan, B. J. DADA2: high-resolution sample inference from Illumina amplicon data. Nat. Methods 13, 581–583 (2016).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Wang, Q., Garrity, G. M., Tiedje, J. M. & Cole, J. R. Naïve Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl. Environ. Microbiol. 73, 5261–5267 (2007).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    McMurdie, P. J. & Holmes, S. phyloseq: an R package for reproducible interactive analysis and graphics of microbiome census data. PLoS ONE 8, e61217 (2013).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Davis, N. M., Proctor, D. M., Holmes, S. P., Relman, D. A. & Callahan, B. J. Simple statistical identification and removal of contaminant sequences in marker-gene and metagenomics data. Microbiome 6, 226 (2018).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Wickham, H. ggplot2: Elegant Graphics for Data Analysis (Springer-Verlag, 2016).Love, M. I., Huber, W. & Anders, S. Moderate estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 15, 550 (2014).Kurtz, Z. D. Sparse and compositionally robust inference of microbial ecological networks. PLoS Comput. Biol. 11, e1004226 (2015).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Csardi, G. & Nepusz, T. The igraph Software Package for Complex Network Research (InterJournal, 2006).Breiman, L. Random forests. Mach. Learn. 45, 5–32 (2001).Kolde, R. pheatmap: pretty heatmaps. R package version 1.0.12 https://CRAN.R-project.org/package=pheatmap (2015).Lin, H. & Das Peddada, S. Analysis of composition of microbiomes with bias correction. Nat. Commun. 11, 3514 (2020).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Oksanen, J. vegan: Community Ecology Package. R package version 2.5.7 https://CRAN.R-project.org/package=vegan (2020).Rouse, G., Pleijel, F. & Tilic, E. Annelida (Oxford Univ. Press, 2022).Ahmed, M. & Holovachov, O. Twenty years after De Ley and Blaxter—How far did we progress in understanding the phylogeny of the phylum Nematoda? Animals 11, 3479 (2021).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Van Steenkiste, N. W. L., Herbert, E. R. & Leander, B. S. Species diversity in the marine microturbellarian Astrotorhynchus bifidus sensu lato (Platyhelminthes: Rhabdocoela) from the Northeast Pacific Ocean. Mol. Phylogenet. Evol. 120, 259–273 (2018). More

  • in

    Determinants of variability in signature whistles of the Mediterranean common bottlenose dolphin

    Wilkins, M. R., Seddon, N. R. & Safran, R. J. Evolutionary divergence in acoustic signals: causes and consequences. Trends Ecol. Evol. 28, 156–166 (2013).PubMed 
    Article 

    Google Scholar 
    Wei, C. Sound production and propagation in cetacean. In Neuroendocrine Regulation of Animal Vocalization (eds Rosenfeld, C. S. & Hoffmann, F.) 267–291 (Academic Press, 2021).Chapter 

    Google Scholar 
    Nakakara, F. Social functions of cetacean acoustic communication. Fish. Sci. 68, 298–301 (2002).Article 

    Google Scholar 
    Caldwell, M. C. & Caldwell, D. K. Vocalization of naive captive dolphins in small groups. Science 159, 1121–1123 (1968).ADS 
    CAS 
    PubMed 
    Article 

    Google Scholar 
    Caldwell, M. C., Caldwell, D. K. & Tyack, P. L. Review of the signature-whistle-hypothesis for the Atlantic bottlenose dolphin. In The bottlenose dolphin (eds Leatherwood, S. & Reeves, R. R.) 199–234 (Academic Press, 1990).Chapter 

    Google Scholar 
    Ford, J. B. Vocal traditions among resident killer whales (Orcinus orca) in coastal waters of British Columbia. Can. J. Zool. 69, 1454–1483 (1991).Article 

    Google Scholar 
    Weilgart, L. & Whitehead, H. Group-specific dialects and geographical variation in coda repertoire in South Pacific sperm whales. Behav. Ecol. Sociobiol. 40, 277–285 (1997).Article 

    Google Scholar 
    Deeck, V. B., Ford, J. K. B. & Spong, P. Dialect change in resident killer whales: implications for vocal learning and cultural transmission. Anim. Behav. 60, 629–638 (2000).Article 

    Google Scholar 
    Chen, Z. & Wiens, J. J. The origins of acoustic communication in vertebrates. Nat. Commun. 11, 369 (2020).ADS 
    CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Morton, E. S. Sources of selection on avian sounds. Am. Nat. 109, 17–34 (1975).ADS 
    Article 

    Google Scholar 
    Irwin, D. E., Thimgan, M. P. & Irwin, J. H. Call divergence is correlated with geographic and genetic distance in greenish warblers (Phylloscopus trochiloides): A strong role for stochasticity in signal evolution?. J. Evol. Biol. 21, 435–448 (2008).CAS 
    PubMed 
    Article 

    Google Scholar 
    Campbell, P. et al. Geographic variation in the songs of Neotropical singing mice: Testing the relative importance of drift and local adaptation. Evol. 64, 1955–1972 (2010).
    Google Scholar 
    Connor, R. C., Wells, R. S., Mann, J. & Read, A. J. The bottlenose dolphin: Social relationships in a fission-fusion society. In Cetacean societies: Field studies of dolphins and whales (eds Mann, J. et al.) 91–126 (University of Chicago Press, Chicago, 2000).
    Google Scholar 
    Janik, V. M. & Sayigh, L. S. Communication in bottlenose dolphins: 50 years of signature whistle research. J. Comp. Physiol. A https://doi.org/10.1007/s00359-013-0817-7 (2013).Article 

    Google Scholar 
    MacFarlane, N. et al. Signature whistles facilitate reunions and/or advertise identity in Bottlenose Dolphins. JASA 141, 3543 (2017).Article 

    Google Scholar 
    Buckstaff, K. C. Effects of watercraft noise on the acoustic behaviour of bottlenose dolphins, Tursiops truncatus, in Sarasota Bay, Florida. Mar. Mam. Sci. 20, 709–725 (2004).Article 

    Google Scholar 
    Cook, M. L. H., Sayigh, L. S., Blum, J. E. & Wells, R. S. Signature-whistle production in undisturbed free-ranging bottlenose dolphins (Tursiops truncatus). Proc. R. Soc. Lond. B. 271, 1043–1049 (2004).Article 

    Google Scholar 
    Watwood, S. L., Owen, E. C. G., Tyack, P. L. & Wells, R. S. Signature whistle use by temporarily restrained and free-swimming bottlenose dolphins, Tursiops truncatus. Anim. Behav. 69, 1373–1386 (2005).Article 

    Google Scholar 
    Sayigh, L. S., Tyack, P. L., Wells, R. S., Scott, M. D. & Irvine, A. B. Sex difference in signature whistle production of free-ranging bottle-nosed dolphins, Tursiops-truncatus. Beh. Ecol. Soc. 36, 171–177 (1995).Article 

    Google Scholar 
    Tyack, P. L. & Sayigh, L. S. Vocal learning in cetaceans. In Social influences on vocal development (eds Snowdon, C. T. & Hausberger, M.) 208–233 (Cambridge University Press, 1997).Chapter 

    Google Scholar 
    Miksis, J. L., Tyack, P. & Buck, J. R. Captive dolphins, Tursiops truncatus, develop signature whistles that match acoustic features of human-made model sounds. JASA 112, 728–739 (2002).Article 

    Google Scholar 
    Fripp, D. et al. Bottlenose dolphin (Tursiops truncatus) calves appear to model their signature whistles on the signature whistles of community members. Anim. Cogn. 8, 17–26 (2005).PubMed 
    Article 

    Google Scholar 
    Janik, V. M. & Slater, P. J. B. Context-specific use suggests that bottlenose dolphin signature whistles are cohesion calls. Anim. Behav. 56, 829–838 (1998).CAS 
    PubMed 
    Article 

    Google Scholar 
    Sayigh, L. S., Tyack, P. L., Wells, R. S. & Scott, M. D. Signature whistles of free-ranging bottlenose dolphins, Tursiops truncatus: mother offspring comparisons. Behav. Ecol. Sociobiol. 26, 247–260 (1990).Article 

    Google Scholar 
    Watwood, S. L., Tyack, P. L. & Wells, R. S. Whistle sharing in paired male bottlenose dolphins, Tursiops truncatus. Behav. Ecol. Sociobiol. 55, 531–543 (2004).Article 

    Google Scholar 
    Janik, V. M., Dehnhardt, G. & Todt, D. Signature whistle variations in a bottlenosed dolphin, Tursiops truncatus. Behav. Ecol. Sociobiol. 35, 243–248 (1994).Article 

    Google Scholar 
    Esch, H. C., Sayigh, L. S. & Wells, R. S. Quantifying parameters of bottlenose dolphin signature whistles. Mar. Mam. Sci. 24, 976–986 (2009).Article 

    Google Scholar 
    Gridley, T. Geographic and species variation in bottlenose dolphin (Tursiops spp.) signature whistle types. PhD Thesis Biology. University of St Andrews (2011).King, S. L. & Janik, V. M. Bottlenose dolphins can use learned vocal labels to address each other. Proc Natl Acad Sci USA 110, 13216–13221 (2013).ADS 
    CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Kriesell, H., Elwen, S. H., Nastasi, A. & Gridley, T. Identification and characteristics of signature whistles in wild bottlenose dolphins (Tursiops truncatus) from Namibia. PLoS ONE 9, e106317 (2014).ADS 
    PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Luis, A. R., Couchinho, M. N. & dos Santos, M. E. Signature whistles in wild bottlenose dolphins: Long term stability and emission rates. Acta Ethol. https://doi.org/10.1007/s10211-015-0230-z (2015).Article 

    Google Scholar 
    Wang, D. W., Würsig, B. & Evans, W. E. Whistles of bottlenose dolphins: Comparisons among populations. Aquatic Mam. 21, 65–77 (1995).
    Google Scholar 
    May-Collado, L. J. & Wartzok, D. A comparison of bottlenose dolphin whistles in the Atlantic Ocean: Factors promoting whistle variation. J. Mammal. 89, 1229–1240 (2008).Article 

    Google Scholar 
    Papale, E. et al. Acoustic divergence between bottlenose dolphin whistles from the Central-Eastern North Atlantic and Mediterranean Sea. Acta Ethol. 17, 155–165 (2014).Article 

    Google Scholar 
    La Manna, G., Rako-Gospić, N., Manghi, M., Picciulin, M. & Sarà, G. Assessing geographical variation on whistle acoustic structure of three Mediterranean populations of common bottlenose dolphin (Tursiops truncatus). Beh. 154, 583–607 (2017).Article 

    Google Scholar 
    La Manna, G. et al. Whistle variation in Mediterranean common bottlenose dolphin: The role of geographical, anthropogenic, social, and behavioral factors. Ecol. Evol. 00, 1–7 (2020).
    Google Scholar 
    Natoli, A., Birkun, A., Aguilar, A., Lopez, A. & Rus Hoelzel, A. Habitat structure and the dispersal of male and female bottlenose dolphins (Tursiops truncatus) based on microsatellite and mitochon-drial DNA analyses. Proc. R. Soc. Lond. B. 272, 1217–2122 (2005).CAS 

    Google Scholar 
    Richardson, W. J., Greene, C. R., Malme, C. I. & Thomson, D. H. Marine mammals and noise (Academic Press, London, 1995).
    Google Scholar 
    Gnone, G., et al. TursioMed: An international project to assess the conservation status of the bottlenose dolphin in the Mediterranean Sea. Final Report (2019).La Manna, G. & Ronchetti, F. Relazione sul monitoraggio della presenza e distribuzione del tursiope Tursiops truncatus nell’area del nord Sardegna comprendente l’Area Marina Protetta Capo Caccia – Isola Piana. Report AMP, 42 (2018).La Manna, G., Ronchetti, F., Sarà, G., Ruiu, A. & Ceccherelli, G. Common bottlenose dolphin protection and sustainable boating: species distribution modeling for effective coastal planning. Front. Mar. Sci. 7, 542648 (2020).Article 

    Google Scholar 
    Pace, D. S. et al. An integrated approach for cetacean knowledge and conservation in the central Mediterranean Sea using research and social media data sources. Aquat. Conserv. 29, 1302–1323 (2019).Article 

    Google Scholar 
    Pace, D. S. et al. Capitoline Dolphins: Residency patterns and abundance estimate of Tursiops truncatus at the Tiber River Estuary (Mediterranean Sea). Biology 10, 275 (2021).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Pulcini, M., Pace, D. S., La Manna, G., Triossi, F. & Fortuna, C. M. Distribution and abundance estimates of bottlenose dolphins (Tursiops truncatus) around Lampedusa Island (Sicily Channel, Italy). Implications for their management. J. Mar. Biol. Assoc. UK 6, 1175–1184 (2013).
    Google Scholar 
    La Manna, G., Ronchetti, F. & Sarà, G. Predicting common bottlenose dolphin habitat preference to dynamically adapt management measures from a Marine Spatial Planning perspective. Ocean Coast. Manag. 130, 317–327 (2016).Article 

    Google Scholar 
    Santostasi, N. L., Bonizzoni, S., Bearzi, G., Eddy, L. & Gimenez, O. A robust design capture-recapture analysis of abundance, survival and temporary emigration of three odontocete species in the Gulf of Corinth, Greece. PLoS ONE 11, e0166650 (2016).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Bearzi, G., Bonizzoni, S. & Gonzalvo, J. Mid-distance movements of common bottlenose dolphins in the coastal waters of Greece. J. Ethol 29, 369–374 (2011).Article 

    Google Scholar 
    Bearzi, G. et al. Dolphins in a scaled-down Mediterranean: The Gulf of Corinth’s odontocetes. In Adv. Mar. Biol. Vol. 75 (eds NotarbartolodiSciara, G. et al.) 297–331 (Academic Press, 2016).
    Google Scholar 
    Pleslić, G. et al. The abundance of common bottlenose dolphins (Tursiops truncatus) in the former special marine reserve of the Cres-Lošinj Archipelago, Croatia. Aquat. Conserv. 25, 125–137 (2015).Article 

    Google Scholar 
    Rako-Gospić, N. et al. Factor associated variations in the home range of a resident Adriatic common bottlenose dolphin population. Mar. Pol. Bul. 124, 234–244 (2017).Article 
    CAS 

    Google Scholar 
    Janik, V. M., King, S. L., Sayigh, L. S. & Wells, R. S. Identifying signature whistles from recordings of groups of unrestrained bottlenose dolphins (Tursiops truncatus). Mar Mam. Sci 29, 1–14 (2013).Article 

    Google Scholar 
    La Manna, G., Manghi, M., Pavan, G., Lo Mascolo, F. & Sarà, G. Behavioural strategy of common bottlenose dolphins (Tursiops truncatus) in response to different kinds of boats in the waters of Lampedusa Island (Italy). Aquat. Conserv. 23, 745–757 (2013).
    Google Scholar 
    R Core Team. R: A Language and Environment for Statistical Computing (R Foundation for Statistical Computing, 2015).
    Google Scholar 
    Zuur, A. F., Ieno, E. N., Walker, N. J., Saveliev, A. A. & Smith, G. H. Mixed effects models and extensions in ecology with R, 579 (Springer, 2009).MATH 
    Book 

    Google Scholar 
    Garamszegi, L. Z. A simple statistical guide for the analysis of behaviour when data are constrained due to practical or ethical reasons. Anim. Beh. 120, 223–234 (2015).Article 

    Google Scholar 
    Pinheiro, J., Bates, D., DebRoy, S., Sarkar, D., & R Core Team. nlme: Linear and Nonlinear Mixed Effects Models. R package version 3.1–137 (2018).Janik, V. M. Source levels and the estimated active space of bottlenose dolphin (Tursiops truncatus) whistles in the Moray Firth, Scotland. J. Comp. Physiol. A Sens. Neural Behav. Physiol 186, 673–680 (2000).CAS 
    Article 

    Google Scholar 
    Quintana-Rizzo, E., Mann, D. A. & Wells, R. S. Estimated communication range of social sounds used by bottlenose dolphins (Tursiops truncatus). JASA 120, 1671–1683 (2006).Article 

    Google Scholar 
    Sayigh, L. S. Development and function of signature whistles of free ranging bottlenose dolphins, Tursiops truncatus. MIT/WHOI joint program (1992).Janik, V. M., Sayigh, L. S. & Wells, R. S. Signature whistle shape conveys identity information to bottlenose dolphins. PNAS 103, 8293–8297 (2006).ADS 
    CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Papale, E., Gamba, M., Perez-Gil, M., Martin, V. M. & Giacoma, C. Dolphins adjust species-specific frequency parameters to compensate for increasing background noise. PLoS ONE 10, e0121711 (2015).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    La Manna, G., Rako-Gospić, N., Manghi, M. & Ceccherelli, G. Influence of environmental, social and behavioural variables on the whistling of the common bottlenose dolphin (Tursiops truncatus). Behav. Ecol. Sociobiol. 73, 12 (2019).Article 

    Google Scholar 
    Ballard, S. M. & Lee, K. M. The acoustics of marine sediments. JASA 13, 18–18 (2017).
    Google Scholar 
    Smolker, R. & Pepper, J. W. Whistle convergence among allied male bottlenose dolphins (Delphinidae, Tursiops sp). Ethology 105, 595–617 (1999).Article 

    Google Scholar 
    Sayigh, L. S., Esch, H. C., Wells, R. S. & Janik, V. M. Facts about signature whistles of bottlenose dolphins (Tursiops truncatus). Anim. Behav. 74, 1631–1642 (2007).Article 

    Google Scholar 
    Jourdan J., et al. Distribution and abundance of bottlenose dolphin (Tursiops truncatus) along French Provençal coast. In Proceeding of the 30th European Cetacean Society Conference, Madeira (2016).Labach, H. et al. Distribution and abundance of common bottlenose dolphin (Tursiops truncatus) over the French Mediterranean continental shelf. Mar. Mam. Sci. 2021, 1–11 (2021).
    Google Scholar 
    Terranova, F. et al. Signature whistles of the demographic unit of bottlenose dolphins (Tursiops truncatus) inhabiting the Eastern Ligurian Sea: characterisation and comparison with the literature. Eur. Zool. J. 88, 771–781 (2021).Article 

    Google Scholar  More