More stories

  • in

    Metagenomic assembled plasmids of the human microbiome vary across disease cohorts

    Dollive, S. A tool kit for quantifying eukaryotic rRNA gene sequences from human microbiome samples. Genome Biol 13, 60 (2012).Article 

    Google Scholar 
    Pausan, M. R. Exploring the archaeome: Detection of archaeal signatures in the human body. Front. Microbiol 10, 2796 (2019).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Shkoporov, A. N. & Hill, C. Bacteriophages of the human gut: The “known unknown” of the microbiome. Cell Host Microbe 25, 195–209 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Clark, D. P., Pazdernik, N. J. & McGehee, M. R. Plasmids. in Molecular Biology, 712–748 (Elsevier, 2019). https://doi.org/10.1016/B978-0-12-813288-3.00023-9.Meinhardt, F., Schaffrath, R. & Larsen, M. Microbial linear plasmids. Appl. Microbiol. Biotechnol 47, 329–336 (1997).CAS 
    PubMed 
    Article 

    Google Scholar 
    Lacroix, B. & Citovsky, V. Transfer of DNA from bacteria to eukaryotes. MBio 7, 00863–16 (2016).Article 

    Google Scholar 
    Łobocka, M. B. Genome of bacteriophage P1. J. Bacteriol. 186, 7032–7068 (2004).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Roux, S., Enault, F., Hurwitz, B. L. & Sullivan, M. B. VirSorter: Mining viral signal from microbial genomic data. PeerJ 3, e985 (2015).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Spaziante, M., Oliva, A., Ceccarelli, G. & Venditti, M. What are the treatment options for resistant Klebsiella pneumoniae carbapenemase (KPC)-producing bacteria?. Expert Opin. Pharmacother. 21, 1781–1787 (2020).PubMed 
    Article 

    Google Scholar 
    Kopotsa, K., Osei Sekyere, J. & Mbelle, N. M. Plasmid evolution in carbapenemase-producing Enterobacteriaceae: A review. Ann. N. Y. Acad. Sci. 1457, 61–91 (2019).CAS 
    PubMed 
    Article 
    ADS 

    Google Scholar 
    Ogilvie, L. A., Firouzmand, S. & Jones, B. V. Evolutionary, ecological and biotechnological perspectives on plasmids resident in the human gut mobile metagenome. Bioengineered 3, 13–31 (2012).Article 

    Google Scholar 
    Jørgensen, T. S., Xu, Z., Hansen, M. A., Sørensen, S. J. & Hansen, L. H. Hundreds of circular novel plasmids and DNA elements identified in a rat cecum metamobilome. PLoS ONE 9, 87924 (2014).Article 
    ADS 
    CAS 

    Google Scholar 
    Kav, A. B. Insights into the bovine rumen plasmidome. Proc. Natl. Acad. Sci. 109, 5452–5457 (2012).CAS 
    PubMed Central 
    Article 
    ADS 

    Google Scholar 
    Brown Kav, A. Unravelling plasmidome distribution and interaction with its hosting microbiome. Environ. Microbiol. 22, 32–44 (2020).PubMed 
    Article 

    Google Scholar 
    Norman, J. M. et al. Disease-specific alterations in the enteric virome in inflammatory bowel disease. Cell 160, 447–460 (2015).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Krishnamurthy, S. R. & Wang, D. Origins and challenges of viral dark matter. Virus Res. 239, 136–142 (2017).CAS 
    PubMed 
    Article 

    Google Scholar 
    Clooney, A. G. et al. Whole-virome analysis sheds light on viral dark matter in inflammatory bowel disease. Cell Host. Microbe 26, 764-778.e5 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Sutton, T. D. S., Clooney, A. G. & Hill, C. Giant oversights in the human gut virome. Gut 69, 1357–1358 (2020).PubMed 
    Article 

    Google Scholar 
    Zuo, T. Gut mucosal virome alterations in ulcerative colitis. Gut 68, 1169–1179 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Pasolli, E. et al. Extensive unexplored human microbiome diversity revealed by over 150,000 genomes from metagenomes spanning age, geography, and lifestyle. Cell 176, 649-662.e20 (2019).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Tamminen, M., Virta, M., Fani, R. & Fondi, M. Large-scale analysis of plasmid relationships through gene-sharing networks. Mol. Biol. Evol. 29, 1225–1240 (2012).CAS 
    PubMed 
    Article 

    Google Scholar 
    Angelakis, E. et al. Treponema species enrich the gut microbiota of traditional rural populations but are absent from urban individuals. New Microbes New Infect 27, 14–21 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Mackie, R. I. et al. Ecology of uncultivated oscillospira species in the rumen of cattle, sheep, and reindeer as assessed by microscopy and molecular approaches. Appl. Environ. Microbiol. 69, 6808–6815 (2003).CAS 
    PubMed 
    PubMed Central 
    Article 
    ADS 

    Google Scholar 
    Konikoff, T. & Gophna, U. Oscillospira: A central, enigmatic component of the human gut microbiota. Trends Microbiol. 24, 523–524 (2016).CAS 
    PubMed 
    Article 

    Google Scholar 
    Chen, Y. et al. High Oscillospira abundance indicates constipation and low BMI in the Guangdong Gut Microbiome Project. Sci. Rep. 10, (2020).Bushman, F. D. Multi-omic analysis of the interaction between clostridioides difficile infection and pediatric inflammatory bowel disease. Cell Host Microbe 28, 422–433 (2020).CAS 
    PubMed 
    Article 

    Google Scholar 
    Willing, B. P. et al. A pyrosequencing study in twins shows that gastrointestinal microbial profiles vary with inflammatory bowel disease phenotypes. Gastroenterology 139, 1844–1854 (2010).PubMed 
    Article 

    Google Scholar 
    Wills, E. S. et al. Fecal microbial composition of ulcerative colitis and Crohn’s disease patients in remission and subsequent exacerbation. PLoS ONE 9, e90981 (2014).PubMed 
    PubMed Central 
    Article 
    ADS 
    CAS 

    Google Scholar 
    Gevers, D. et al. The treatment-naive microbiome in new-onset Crohn’s disease. Cell Host Microbe 15, 382–392 (2014).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Halfvarson, J. Dynamics of the human gut microbiome in inflammatory bowel disease. Nat. Microbiol. 2, 17004 (2017).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Pascal, V. A microbial signature for Crohn’s disease. Gut 66, 813–822 (2017).CAS 
    PubMed 
    Article 

    Google Scholar 
    Nitzan, O., Elias, M., Chazan, B., Raz, R. & Saliba, W. Clostridium difficile and inflammatory bowel disease: Role in pathogenesis and implications in treatment. World J. Gastroenterol. 19, 7577–7585 (2013).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Clayton, E. M. et al. The vexed relationship between Clostridium difficile and inflammatory bowel disease: an assessment of carriage in an outpatient setting among patients in remission. Am. J. Gastroenterol. 104, 1162–1169 (2009).PubMed 
    Article 
    ADS 

    Google Scholar 
    Tariq, R. et al. Efficacy of fecal microbiota transplantation for recurrent C.Marcella, C. Systematic review: The global incidence of faecal microbiota transplantation-related adverse events from 2000 to 2020. Aliment. Pharmacol. Ther. https://doi.org/10.1111/apt.16148 (2020).Article 
    PubMed 

    Google Scholar 
    Shkoporov, A. N. et al. The human gut virome is highly diverse, stable, and individual specific. Cell Host Microbe 26, 527-541.e5 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Bolger, A. M., Lohse, M. & Usadel, B. Trimmomatic: A flexible trimmer for Illumina sequence data. Bioinformatics 30, 2114–2120 (2014).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Fraser-Liggett, C. Metagenomic analysis of the structure and function of the human gut microbiota in Crohn’s disease. Nat. Preced. [Internet] (2010).Barton, W. et al. The microbiome of professional athletes differs from that of more sedentary subjects in composition and particularly at the functional metabolic level. Gut (2017).Mira-Pascual, L. Microbial mucosal colonic shifts associated with the development of colorectal cancer reveal the presence of different bacterial and archaeal biomarkers. J. Gastroenterol. 50, 167–179 (2015).CAS 
    PubMed 
    Article 
    ADS 

    Google Scholar 
    Rampelli, S. Shotgun metagenomics of gut microbiota in humans with up to extreme longevity and the increasing role of xenobiotic degradation. mSystems 5, (2020).Monaghan, T. M. Metagenomics reveals impact of geography and acute diarrheal disease on the Central Indian human gut microbiome. Gut Microbes 12, 1752605 (2020).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Chu, D. M. Maturation of the infant microbiome community structure and function across multiple body sites and in relation to mode of delivery. Nat. Med. 23, 314–326 (2017).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    MD, D. G., K, F., C, C. & EL, C. Whole genome metagenomic analysis of the gut microbiome of differently fed infants identifies differences in microbial composition and functional genes, including an absent CRISPR/Cas9 gene in the formula-fed cohort. Hum. Microbiome J. 12, (2019).Qian, Y. et al. Gut metagenomics-derived genes as potential biomarkers of Parkinson’s disease. Brain J. Neurol. 143, 2474–2489 (2020).Article 

    Google Scholar 
    Kao, D. Effect of oral capsule- vs colonoscopy-delivered fecal microbiota transplantation on recurrent clostridium difficile infection: A randomized clinical trial. JAMA 318, 1985–1993 (2017).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Nurk, S., Meleshko, D., Korobeynikov, A. & Pevzner, P. A. metaSPAdes: A new versatile metagenomic assembler. Genome Res. 27, 824–834 (2017).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Hyatt, D. et al. Prodigal: prokaryotic gene recognition and translation initiation site identification. BMC Bioinform. 11, 119 (2010).Article 
    CAS 

    Google Scholar 
    Finn, R. D., Clements, J. & Eddy, S. R. HMMER web server: Interactive sequence similarity searching. Nucleic Acids Res. 39, W29–W37 (2011).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Guerin, E. et al. Biology and taxonomy of crAss-like bacteriophages, the most abundant virus in the human gut. (2018). https://doi.org/10.1101/295642.Grazziotin, A. L., Koonin, E. V. & Kristensen, D. M. Prokaryotic Virus Orthologous Groups (pVOGs): A resource for comparative genomics and protein family annotation. Nucleic Acids Res. 45, D491–D498 (2017).CAS 
    PubMed 
    Article 

    Google Scholar 
    Langmead, B. & Salzberg, S. L. Fast gapped-read alignment with Bowtie 2. Nat. Methods 9, 357–359 (2012).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Li, H. et al. The sequence alignment/map format and SAMtools. Bioinformatics 25, 2078–2079 (2009).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Quinlan, A. R. & Hall, I. M. BEDTools: A flexible suite of utilities for comparing genomic features. Bioinformatics 26, 841–842 (2010).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Edgar, R. C. PILER-CR: Fast and accurate identification of CRISPR repeats. BMC Bioinform. 8, 18 (2007).Article 
    CAS 

    Google Scholar 
    R Core Team. R: A Language and Environment for Statistical Computing. https://www.R-project.org/. (2019). Accessed Aug 2021–Mar 2022.Wickham, H. Reshaping Data with the reshape Package. J. Stat. Softw. 21, 1–20 (2007).Article 

    Google Scholar 
    Jari Oksanen et al. vegan: Community Ecology Package. (2019).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 
    ADS 

    Google Scholar 
    Kassambara, A. ggpubr: ‘ggplot2’ based publication ready plots. (2019).Wickham, H. ggplot2: Elegant Graphics for Data Analysis (Springer-Verlag, 2016).MATH 
    Book 

    Google Scholar 
    Gu, Z., Gu, L., Eils, R., Schlesner, M. & Brors, B. Circlize implements and enhances circular visualization in R. Bioinformatics 30, 2811–2812 (2014).CAS 
    PubMed 
    Article 

    Google Scholar 
    Flor M. chorddiag: Interactive Chord Diagrams [Internet]. (2020).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 
    Hulsen, T., Vlieg, J. & Alkema, W. BioVenn—A web application for the comparison and visualization of biological lists using area-proportional Venn diagrams. BMC Genom. 9, (2008).Stothard, P. & Wishart, D. S. Circular genome visualization and exploration using CGView. Bioinform. Oxf. Engl. 21, 537–539 (2005).CAS 
    Article 

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

    Google Scholar 
    Huerta-Cepas, J. et al. eggNOG 4.5: A hierarchical orthology framework with improved functional annotations for eukaryotic, prokaryotic and viral sequences. Nucleic Acids Res. 44, D286–D293 (2016).CAS 
    PubMed 
    Article 

    Google Scholar 
    McArthur, A. G. et al. The comprehensive antibiotic resistance database. Antimicrob. Agents Chemother. 57, 3348–3357 (2013).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Li, W. & Godzik, A. Cd-hit: A fast program for clustering and comparing large sets of protein or nucleotide sequences. Bioinformatics 22, 1658–1659 (2006).CAS 
    PubMed 
    Article 

    Google Scholar  More

  • in

    Dental macrowear reveals ecological diversity of Gorilla spp.

    Fossey, D. & Harcourt, D. H. Feeding ecology of free-ranging mountain gorilla (Gorilla gorilla beringei). In Primate ecology (ed. Clutton-Brock, T. H.) 415–447 (Academy Press, New York, 1977).Watts, D. P. Composition and variability of mountain gorilla diets in the central Virungas. Am. J. Primatol. 7, 323–356 (1984).PubMed 
    Article 

    Google Scholar 
    Watts, D. Comparative socio-ecology of gorillas. In Great Ape Societies (eds. McGrew, W. C., Marchant, L. F. & Nishida, T.) 16–28 (Cambridge University Press, Cambridge, 1986).Doran, D. M. & McNeilage, A. Gorilla ecology and behavior. Evol. Anthropol. 6, 120–131 (1988).Article 

    Google Scholar 
    Xue, Y. et al. Mountain gorilla genomes reveal the impact of long-term population decline and inbreeding. Science 348, 242–245 (2015).ADS 
    CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Mittermeier, R. A., Rylands, A. B. & Wilson, D. E. Handbook of the mammals of the world. Primates Vol. 3 (Lynx Edicions, 2013).
    Google Scholar 
    Groves, C. Primate Taxonomy (Smithsonian Institution Press, 2001).
    Google Scholar 
    Cooksey, K. E. & Morgan, D. B. Gorilla (Gorilla). In The International encyclopedia of primatology, Vol. 1 (ed. Fuentes, A.) 472–477 (Wiley, Hoboken, 2017).McFarland, K. L. Ecology of cross river gorillas (Gorilla gorilla diehli) on Afi mountain, Cross River State, Nigeria. Ph.D. Dissertation. City University of New York, USA (2007).Rogers, M. E., Maisels, F., Wiliamson, E. A., Fernandez, M. & Tutin, C. E. G. Gorilla diet in the Lopé Reserve, Gabon: a nutritional analysis. Oecologia 84, 326–339 (1990).ADS 
    Article 

    Google Scholar 
    van Casteren, A., Wright, E., Kupczik, K. & Robbins, M. M. Unexpected hard-object feeding in Western lowland gorillas. Am. J. Phys. Anthropol. 170, 433–438 (2019).PubMed 
    Article 

    Google Scholar 
    Masi, S., Cipolletta, C. & Robbins, M. M. Western lowland gorillas (Gorilla gorilla gorilla) change their activity patterns in response to frugivory. Am. J. Primatol. 71, 91–100 (2009).PubMed 
    Article 

    Google Scholar 
    Yamagiwa, J., Basabose, A. K., Kaleme, K. & Yumoto, T. Diet of grauer’s gorillas in montane forest of Kahuzi, Democratic Republic of Congo. Int. J. Primatol. 26, 1345–1373 (2005).Article 

    Google Scholar 
    Grueter, C. C. et al. Long-term temporal and spatial dynamics of food availability for endangered mountain gorillas in Volcanoes National Park, Rwanda. Am. J. Primatol. 75, 267–280 (2013).PubMed 
    Article 

    Google Scholar 
    Ostrofsky, K. R. & Robbins, M. M. Fruit-feeding and activity patterns of mountain gorillas (Gorilla beringei beringei) in Bwindi Impenetrable National Park, Uganda. Am. J. Phys. Anthropol. 173, 3–20 (2020).PubMed 
    Article 

    Google Scholar 
    Berthaume, M. A. Tooth cusp sharpness as a dietary correlate in great apes. Am. J. Phys. Anthropol. 153, 226–235 (2014).PubMed 
    Article 

    Google Scholar 
    King, S. J. et al. Dental senescence in a long-lived primate links infant survival to rainfall. Proc. Natl. Acad. Sci. USA 102, 16579–16583 (2005).ADS 
    CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Berthaume, M. A. & Schroer, K. Extant ape dental topography and its implications for reconstructing the emergence of early Homo. J. Hum. Evol. 112, 15–29 (2017).PubMed 
    Article 

    Google Scholar 
    Sheine, W. S. & Kay, R. F. An analysis of chewed food particle size and its relationship to molar structure in the primates Cheirogaleus medius and Galago senegalensis and the insectivoran Tupaia glis. Am. J. Phys. Anthropol. 47, 15–20 (1977).Article 

    Google Scholar 
    Galbany, J., Estebaranz, F., Martínez, L. M. & Pérez-Pérez, A. Buccal dental microwear variability in extant African Hominoidea: taxonomy versus ecology. Primates 50, 221–230 (2009).PubMed 
    Article 

    Google Scholar 
    Scott, R. S., Teaford, M. F. & Ungar, P. S. Dental microwear texture and anthropoid diets. Am. J. Phys. Anthropol. 147, 551–579 (2012).PubMed 
    Article 

    Google Scholar 
    Teaford, M. F. & Oyen, O. J. In vivo and in vitro turnover in dental microwear. Am. J. Phys. Anhtropol. 80, 447–460 (1989).CAS 
    Article 

    Google Scholar 
    Stuhlträger, J. et al. Dental wear patterns reveal dietary ecology and season of death in a historical chimpanzee population. PLoS ONE 16, e0251309 (2021).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Elgart, A. A. Dental wear, wear rate, and dental disease in the African apes. Am. J. Primatol. 72, 481–491 (2010).PubMed 

    Google Scholar 
    Berthaume, M. A. Food mechanical properties and dietary ecology. Am. J. Phys. Anhtropol. 159, 79–104 (2016).Article 

    Google Scholar 
    Galbany, J. et al. Tooth wear and feeding ecology in mountain gorillas from Volcanoes National Park, Rwanda. Am. J. Phys. Anhtropol. 159, 457–465 (2016).Article 

    Google Scholar 
    Janis, C. M. The correlation between diet and dental wear in herbivorous mammals, and its relationship to the determination of diets of extinct species, in Evolutionary paleobiology of behavior and coevolution (ed. Boucot, A. J.) 241–259 (Elsevier, Amsterdam, 1990).Knight-Sadler, J. & Fiorenza, L. Tooth wear inclination in great ape molars. Folia Primatol. 88, 223–236 (2017).Article 

    Google Scholar 
    Kullmer, O. et al. Technical note: Occlusal fingerprint analysis: Quantification of tooth wear pattern. Am. J. Phys. Anthropol. 139, 600–605 (2009).PubMed 
    Article 

    Google Scholar 
    Fiorenza, L. et al. Molar macrowear reveals Neanderthal eco-geographical dietary variation. PLoS ONE 6, e14769 (2011).ADS 
    CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Fiorenza, L., Benazzi, S., Oxilia, G. & Kullmer, O. Functional relationship between dental macrowear and diet in Late Pleistocene and recent modern human populations. Int. J. Osteoarchaeol. 28, 153–161 (2018).Article 

    Google Scholar 
    Fiorenza, L. et al. The functional role of the Carabelli trait in early and late hominins. J. Hum. Evol. 145, 102816 (2020).PubMed 
    Article 

    Google Scholar 
    Fiorenza, L. & Kullmer, O. Occlusion in an adult male gorilla with a supernumerary maxillary premolar. Int. J. Primatol. 37, 762–777 (2016).Article 

    Google Scholar 
    Kullmer, O., Menz, U., & Fiorenza, L. Occlusal fingerprint analysis (OFA) reveal dental occlusal behaviour in primate teeth. In T. Martin & W. von Koenigswald (Eds.), T Martin, W von Koenigswald, K-H Südekum), Mammalian teeth: form and function. (pp. 25–43). Munich, Germany: Dr. F. Pfeil (2020)Stuhlträger, J. et al. Dental wear patterns reveal dietary ecology and season of death in a historical chimpanzee population. PLoS ONE 16, e0251309 (2021).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Fiorenza, L., Benazzi, S., Estalrrich, A., & Kullmer, O. Diet and cultural diversity in Neanderthals and modern humans from dental macrowear analyses. In C. Schmidt & J. T. Watson (Eds.), Dental wear in evolutionary and biocultural contexts (pp. 39–72). London, UK: Academic Press (2020).M’Kirera, F. & Ungar, P. S. Occlusal relief changes with molar wear in Pan troglodytes troglodytes and Gorilla gorilla gorilla. Am. J. Primatol. 60, 31–41 (2003).PubMed 
    Article 

    Google Scholar 
    Galbany, J. et al. Age-related tooth wear differs between forest and savanna primates. PLoS ONE 9, e94938 (2014).ADS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Leigh, S. R. & Shea, B. T. Ontogeny and the evolution of adult body size dimorphism in apes. Am. J. Primatol. 36, 37–60 (1995).PubMed 
    Article 

    Google Scholar 
    Watts, D. P. Environmental influences on mountain gorilla time budgets. Am. J. Primatol. 15, 195–211 (1988).PubMed 
    Article 

    Google Scholar 
    Doran, D. M. et al. Western lowland gorilla diet and resource availability: New evidence, cross-site comparisons, and reflections on indirect sampling methods. Am. J. Primatol. 58, 91–116 (2002).PubMed 
    Article 

    Google Scholar 
    Zanolli, C. et al. Evidence of increased hominid diversity in the Early and Middle Pleistocene of Indonesia. Nat. Ecol. Evol. 3, 755–764 (2019).PubMed 
    Article 

    Google Scholar 
    Krueger, K. L., Scott, J. R., Kay, R. F. & Ungar, P. S. Dental microwear textures of “phase I” and “phase II” facets. Am. J. Phys. Anthropol. 137, 485–490 (2008).PubMed 
    Article 

    Google Scholar 
    Kay, R. F. & Hiiemae, K. M. Jaw movement and tooth use in recent and fossil primates. Am. J. Phys. Anthropol. 40, 227–256 (1974).CAS 
    PubMed 
    Article 

    Google Scholar 
    Wall, C. E., Vinyard, C. J., Johnson, K. R., Williams, S. H. & Hylander, W. L. Phase II jaw movements and masseter muscle activity during chewing in Papio anubis. Am. J. Phys. Anthropol. 129, 215–224 (2006).PubMed 
    Article 

    Google Scholar 
    Glowacka, H. et al. Toughness of the Virunga mountain gorilla (Gorilla beringei beringei) diet across an altitudinal gradient. Am. J. Primatol. 79, e22661 (2017).Article 

    Google Scholar 
    Cooper, J. E. & Hull, G. Gorilla pathology and health (Academic Press, 2017).
    Google Scholar 
    Hammerton, R., Hunt, K. A. & Riley, L. M. An investigation into keeper opinions of great apes diet and abnormal behaviour. J. Zoo Aquar. Res. 7, 170–178 (2019).
    Google Scholar 
    Kay, R. F. Mastication, molar tooth structure and diet in primates. Ph.D. thesis, Yale University, New Haven, CT (1973).Smith, B. H. Patterns of molar wear in hunter-gatherers and agriculturalists. Am. J. Phys. Anhtropol. 63, 39–56 (1984).CAS 
    Article 

    Google Scholar 
    Maier, W. & Schneck, G. Konstruktionsmorphologische Untersuchungen am Gebiß der hominoiden Primaten. Z. Morphol. Anthropol. 72, 127–169 (1981).CAS 
    PubMed 
    Article 

    Google Scholar 
    Fiorenza, L., Benazzi, S., Tausch, J., Kullmer, O. & Schrenk, F. Identification reassessment of the isolated tooth Krapina D58 through Occlusal Fingerprint Analysis. Am. J. Phys. Anthropol. 143, 306–312 (2010).PubMed 
    Article 

    Google Scholar 
    Kullmer, O., Huck, M., Engel, K., Schrenk, F. & Bromage, T. Hominid Tooth Pattern Database (HOTPAD) derived from optical 3D topometry. In Three-dimensional imaging in paleoanthropology and prehistoric archaeology (eds. Mafart, B. & Delingette, H.) 71–82 (Acts of the XIVth UISPP Congress, BAR Int. Ser.1049, 2002).Hammer, Ø. & Harper, D. Paleontological data analysis (Blackwell Publishing, 2006).
    Google Scholar 
    Brown, M. B. & Forsythe, A. B. Robust tests for the equality of variances. J. Am. Stat. Assoc. 69, 364–367 (1974).MATH 
    Article 

    Google Scholar 
    Noguchi, K. & Gel, Y. R. Combination of Levene-type tests and a finite-intersection method for testing equality of variances against ordered alternatives. J. Nonparam. Stat. 22, 897–913 (2010).MathSciNet 
    MATH 
    Article 

    Google Scholar 
    Gastwirth, J. L., et al. Lawstat: tools for biostatistics, public policy, and law. R package version 3.4. https://CRAN.R-project.org/package=lawstat (2020).R Core Team. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. https://www.R-project.org/. (2021)Oksanen, J. et al. Vegan: Community Ecology Package. R package version 2.5–7. https://CRAN.R-project.org/package=vegan (2020).Martinez Arbizu, P. PairwiseAdonis: pairwise multilevel comparison using adonis. R package version 0.4 (2017).Hammer, Ø., Harper, D. A. T. & Ryan, P. D. PAST: Palaeontological statistics software package for education and data analysis. Palaeontol. Electron. 4, 9 (2001).
    Google Scholar  More

  • in

    Characterization of intestinal microbiota in normal weight and overweight Border Collie and Labrador Retriever dogs

    Lund, E. M., Armstrong, P. J., Kirk, C. A. & Klausner, J. S. Prevalence and risk factors for obesity in adult dogs from private US veterinary practices. Int. J. Appl. Res. Vet. Med. 4(2), 177 (2006).
    Google Scholar 
    German, A. J. The growing problem of obesity in dogs and cats. J. Nutr. 136(7), 1940S-1946S (2006).CAS 
    PubMed 
    Article 

    Google Scholar 
    Courcier, E. A., Thomson, R. M., Mellor, D. J. & Yam, P. S. An epidemiological study of environmental factors associated with canine obesity. J. Small Anim. Pract. 51(7), 362–367 (2010).CAS 
    PubMed 
    Article 

    Google Scholar 
    Mao, J., Xia, Z., Chen, J. & Yu, J. Prevalence and risk factors for canine obesity surveyed in veterinary practices in Beijing, China. Prev. Vet. Med. 112(3–4), 438–442 (2013).PubMed 
    Article 

    Google Scholar 
    Payan-Carreira, R., Sargo, T. & Nascimento, M. M. Canine obesity in Portugal: Perceptions on occurrence and treatment determinants. Acta Vet. Scand. 57(1), 1–1 (2015).Article 

    Google Scholar 
    Chandler, M. et al. Obesity and associated comorbidities in people and companion animals: A one health perspective. J. Comp. Pathol. 156(4), 296–309 (2017).CAS 
    PubMed 
    Article 

    Google Scholar 
    Montoya-Alonso, J. A. et al. Prevalence of canine obesity, obesity-related metabolic dysfunction, and relationship with owner obesity in an obesogenic region of Spain. Front. Vet. Sci. 4, 59 (2017).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Muñoz-Prieto, A. et al. European dog owner perceptions of obesity and factors associated with human and canine obesity. Sci. Rep. 8(1), 1–10 (2018).Article 
    CAS 

    Google Scholar 
    Marshall, W. G., Bockstahler, B. A., Hulse, D. A. & Carmichael, S. A review of osteoarthritis and obesity: Current understanding of the relationship and benefit of obesity treatment and prevention in the dog. Vet. Comp. Orthop. Traumatol. 22(05), 339–345 (2009).CAS 
    PubMed 
    Article 

    Google Scholar 
    Zoran, D. L. Obesity in dogs and cats: A metabolic and endocrine disorder. Vet. Clin. N. Am. Small Anim. Pract. 40(2), 221–239 (2010).Article 

    Google Scholar 
    Tvarijonaviciute, A. et al. Obesity-related metabolic dysfunction in dogs: A comparison with human metabolic syndrome. BMC Vet. Res. 8(1), 147 (2012).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Hoenig, M. Comparative aspects of human, canine, and feline obesity and factors predicting progression to diabetes. Vet. Sci. 1(2), 121–135 (2014).Article 

    Google Scholar 
    Yam, P. S. et al. Impact of canine overweight and obesity on health-related quality of life. Prev. Vet. Med. 127, 64–69 (2016).CAS 
    PubMed 
    Article 

    Google Scholar 
    Sandøe, P., Palmer, C., Corr, S., Astrup, A. & Bjørnvad, C. R. Canine and feline obesity: A One Health perspective. Vet. Rec. 175(24), 610–616 (2014).PubMed 
    Article 

    Google Scholar 
    Salt, C., Morris, P. J., Wilson, D., Lund, E. M. & German, A. J. Association between life span and body condition in neutered client-owned dogs. J. Vet. Intern. Med. 33(1), 89–99 (2019).PubMed 

    Google Scholar 
    Switonski, M. & Mankowska, M. Dog obesity—The need for identifying predisposing genetic markers. Res. Vet. Sci. 95(3), 831–836 (2013).CAS 
    PubMed 
    Article 

    Google Scholar 
    Mankowska, M. et al. Sequence analysis of three canine adipokine genes revealed an association between TNF polymorphisms and obesity in Labrador dogs. Anim. Genet. 47(2), 245–249 (2016).CAS 
    PubMed 
    Article 

    Google Scholar 
    Raffan, E. et al. A deletion in the canine POMC gene is associated with weight and appetite in obesity-prone Labrador retriever dogs. Cell Metab. 23(5), 893–900 (2016).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Suchodolski, J. S. Intestinal microbiota of dogs and cats: A bigger world than we thought. Anim. Pract. 41(2), 261–272 (2011).
    Google Scholar 
    Barko, P. C., McMichael, M. A., Swanson, K. S. & Williams, D. A. The gastrointestinal microbiome: A review. J. Vet. Intern. Med. 32(1), 9–25 (2018).CAS 
    PubMed 
    Article 

    Google Scholar 
    Turnbaugh, P. J. et al. An obesity-associated gut microbiome with increased capacity for energy harvest. Nature 444(7122), 1027–1031 (2006).PubMed 
    Article 
    ADS 

    Google Scholar 
    Bäckhed, F. et al. The gut microbiota as an environmental factor that regulates fat storage. Proc. Natl. Acad. Sci. 101(44), 15718–15723 (2004).PubMed 
    PubMed Central 
    Article 
    ADS 
    CAS 

    Google Scholar 
    Ghazalpour, A., Cespedes, I., Bennett, B. J. & Allayee, H. Expanding role of gut microbiota in lipid metabolism. Curr. Opin. Lipidol. 27(2), 141 (2016).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Losasso, C. et al. Assessing the influence of vegan, vegetarian and omnivore oriented westernized dietary styles on human gut microbiota: A cross sectional study. Front. Microbiol. 9, 317 (2018).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Pizarroso, N. A., Fuciños, P., Gonçalves, C., Pastrana, L. & Amado, I. R. A Review on the role of food-derived bioactive molecules and the microbiota—Gut–brain axis in satiety regulation. Nutrients 13(2), 632. https://doi.org/10.3390/nu13020632 (2021).CAS 
    Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Boulangé, C. L., Neves, A. L., Chilloux, J., Nicholson, J. K. & Dumas, M. E. Impact of the gut microbiota on inflammation, obesity, and metabolic disease. Genome Med. 8(1), 1–12 (2016).Article 
    CAS 

    Google Scholar 
    Ley, R. E. et al. Obesity alters gut microbial ecology. Proc. Natl. Acad. Sci. USA 102(31), 11070–11075 (2005).CAS 
    PubMed 
    PubMed Central 
    Article 
    ADS 

    Google Scholar 
    Ley, R. E., Turnbaugh, P. J., Klein, S. & Gordon, J. I. Human gut microbes associated with obesity. Nature 444(7122), 1022–1023 (2006).CAS 
    PubMed 
    Article 
    ADS 

    Google Scholar 
    Zhi, C. et al. Connection between gut microbiome and the development of obesity. Eur. J. Clin. Microbiol. Infect. Dis. 38(11), 1987–1998 (2019).PubMed 
    Article 

    Google Scholar 
    Huang, Z., Pan, Z., Yang, R., Bi, Y. & Xiong, X. The canine gastrointestinal microbiota: Early studies and research frontiers. Gut Microbes 11(4), 635–654 (2020).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Swanson, K. S. et al. Phylogenetic and gene-centric metagenomics of the canine intestinal microbiome reveals similarities with humans and mice. ISME J. 5(4), 639–649 (2011).CAS 
    PubMed 
    Article 

    Google Scholar 
    Coelho, L. P. et al. Similarity of the dog and human gut microbiomes in gene content and response to diet. Microbiome 6(1), 1–11 (2018).Article 

    Google Scholar 
    Hand, D., Wallis, C., Colyer, A. & Penn, C. W. Pyrosequencing the canine faecal microbiota: Breadth and depth of biodiversity. PLoS ONE 8(1), e53115 (2013).CAS 
    PubMed 
    PubMed Central 
    Article 
    ADS 

    Google Scholar 
    Handl, S. et al. Faecal microbiota in lean and obese dogs. FEMS Microbiol. Ecol. 84(2), 332–343 (2013).CAS 
    PubMed 
    Article 

    Google Scholar 
    Park, H. J. et al. Association of obesity with serum leptin, adiponectin, and serotonin and gut microflora in beagle dogs. J. Vet. Intern. Med. 29(1), 43–50 (2015).PubMed 
    Article 

    Google Scholar 
    Park, H. J. et al. Fecal microbiota analysis of obese dogs with underlying diseases: A pilot study. Korean J. Vet. Res. 55(3), 205–208 (2015).Article 

    Google Scholar 
    Beloshapka, A. N., Forster, G. M., Holscher, H. D., Swanson, K. S. & Ryan, E. P. Fecal microbial communities of overweight and obese client-owned dogs fed cooked bean powders as assessed by 454-pyrosequencing. J. Vet. Sci. Technol. 7(366), 2 (2016).
    Google Scholar 
    Li, Q., Lauber, C. L., Czarnecki-Maulden, G., Pan, Y. & Hannah, S. S. Effects of the dietary protein and carbohydrate ratio on gut microbiomes in dogs of different body conditions. MBio 8(1), e01703-e1716 (2017).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Kieler, I. N. et al. Gut microbiota composition may relate to weight loss rate in obese pet dogs. Vet. Med. Sci. 3(4), 252–262 (2017).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Forster, G. M. et al. A comparative study of serum biochemistry, metabolome and microbiome parameters of clinically healthy, normal weight, overweight, and obese companion dogs. Top. Companion Anim. Med. 33(4), 126–135 (2018).PubMed 
    Article 

    Google Scholar 
    Salas-Mani, A. et al. Fecal microbiota composition changes after a BW loss diet in beagle dogs. J. Anim. Sci. 96(8), 3102–3111 (2018).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Alexander, C. et al. Effects of prebiotic inulin-type fructans on blood metabolite and hormone concentrations and faecal microbiota and metabolites in overweight dogs. Br. J. Nutr. 120(6), 711–720 (2018).CAS 
    PubMed 
    Article 

    Google Scholar 
    Herstad, K. M. et al. A diet change from dry food to beef induces reversible changes on the faecal microbiota in healthy, adult client-owned dogs. BMC Vet. Res. 13(1), 147 (2017).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Kim, Y. S., Unno, T., Kim, B. Y. & Park, M. S. Sex differences in gut microbiota. World J. Mens Health 38(1), 48–60 (2020).PubMed 
    Article 

    Google Scholar 
    Xu, J. et al. The response of canine faecal microbiota to increased dietary protein is influenced by body condition. BMC Vet. Res. 13(1), 374 (2017).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Masuoka, H. et al. Transition of the intestinal microbiota of dogs with age. PLoS ONE 12, e0181739 (2016).Article 
    CAS 

    Google Scholar 
    Mizukami, K. et al. Age-related analysis of the gut microbiome in a purebred dog colony. FEMS Microbiol. Lett. 366(8), fnz095 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Alessandri, G. et al. Metagenomic dissection of the canine gut microbiota: Insights into taxonomic, metabolic and nutritional features. Environ. Microbiol. 21(4), 1331–1343 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Xu, H. et al. Oral administration of compound probiotics improved canine feed intake, weight gain, immunity and intestinal microbiota. Front. Immunol. 10, 666 (2019).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Reddy, K. E. et al. Impact of breed on the fecal microbiome of dogs under the same dietary condition. J. Microbiol. Biotechnol. 29(12), 1947–1956 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    O’Neill, D. G., Church, D. B., McGreevy, P. D., Thomson, P. C. & Brodbelt, D. C. Prevalence of disorders recorded in dogs attending primary-care veterinary practices in England. PLoS ONE 9(3), e90501 (2014).PubMed Central 
    Article 
    ADS 

    Google Scholar 
    Vilson, Å. et al. Disentangling factors that shape the gut microbiota in German Shepherd dogs. PLoS ONE 13(3), e0193507 (2018).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Song, S. J. et al. Cohabiting family members share microbiota with one another and with their dogs. Elife 2, e00458 (2013).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Guard, B. C. et al. Characterization of the fecal microbiome during neonatal and early pediatric development in puppies. PLoS ONE 12(4), e0175718 (2017).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Greer, K. A., Canterberry, S. C. & Murphy, K. E. Statistical analysis regarding the effects of height and weight on life span of the domestic dog. Res. Vet. Sci. 82(2), 208–214 (2007).PubMed 
    Article 

    Google Scholar 
    Fleming, J. M., Creevy, K. E. & Promislow, D. E. L. Mortality in North American dogs from 1984 to 2004: An investigation into age-, size-, and breed-related causes of death. J. Vet. Int. Med. 25(2), 187–198 (2011).CAS 
    Article 

    Google Scholar 
    Oberbauer, A. M., Belanger, J. & Famula, T. R. A review of the impact of neuter status on expression of inherited conditions in dogs. Front. Vet. Sci. 6, 397 (2019).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Pilla, R. & Suchodolski, J. S. The role of the canine gut microbiome and metabolome in health and gastrointestinal disease. Front. Vet. Sci. 6, 498 (2020).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Bermingham, E. N., Maclean, P., Thomas, D. G., Cave, N. J. & Young, W. Key bacterial families (Clostridiaceae, Erysipelotrichaceae and Bacteroidaceae) are related to the digestion of protein and energy in dogs. PeerJ 5, e3019 (2017).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Kim, J., An, J. U., Kim, W., Lee, S. & Cho, S. Differences in the gut microbiota of dogs (Canis lupus familiaris) fed a natural diet or a commercial feed revealed by the Illumina MiSeq platform. Gut Pathog. 9, 68–68 (2017).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Mori, A. et al. Comparison of the effects of four commercially available prescription diet regimens on the fecal microbiome in healthy dogs. J. Vet. Med. Sci. 81, 1783–1790 (2019).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Apper, E. et al. Relationships between gut microbiota, metabolome, body weight, and glucose homeostasis of obese dogs fed with diets differing in prebiotic and protein content. Microorganisms 8(4), 513 (2020).CAS 
    PubMed Central 
    Article 

    Google Scholar 
    Wernimont, S. M. et al. The effects of nutrition on the gastrointestinal microbiome of cats and dogs: Impact on health and disease. Front. Microbiol. https://doi.org/10.3389/fmicb.2020.01266 (2020).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Schauf, S. et al. Effect of dietary fat to starch content on fecal microbiota composition and activity in dogs. J. Anim. Sci. 96(9), 3684–3698 (2018).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Bresciani, F. et al. Effect of an extruded animal protein-free diet on fecal microbiota of dogs with food-responsive enteropathy. J. Vet. Intern. Med. 32(6), 1903–1910 (2018).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Madsen, L., Myrmel, L. S., Fjære, E., Liaset, B. & Kristiansen, K. Links between dietary protein sources, the gut microbiota, and obesity. Front. Physiol. 8, 1047 (2017).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    EU law and publications. Regulation (EC) No 767/2009 of the European parliament and of the council of 13 July 2009 on the placing on the market and use of feed, amending European Parliament and council regulation (EC) No 1831/2003 and repealing council directive 79/373/EEC, commission directive 80/511/EEC, council directives 82/471/EEC, 83/228/EEC, 93/74/EEC, 93/113/EC and 96/25/EC and commission decision 2004/217/EC. OJEC L229, 1–28 (2009).
    Google Scholar 
    Paßlack, N. et al. Impact of the dietary inclusion of dried food residues on the apparent nutrient digestibility and the intestinal microbiota of dogs. Arch. Anim. Nutr. 75(4), 311–327 (2021).PubMed 
    Article 

    Google Scholar 
    Macedo, H. T. et al. Weight-loss in obese dogs promotes important shifts in fecal microbiota profile to the extent of resembling microbiota of lean dogs. Anim. Microbiome 4(1), 1–13 (2022).Article 
    CAS 

    Google Scholar 
    Zhang, X. et al. Human gut microbiota changes reveal the progression of glucose intolerance. PLoS ONE 8(8), e71108 (2013).CAS 
    PubMed 
    PubMed Central 
    Article 
    ADS 

    Google Scholar 
    Remely, M. et al. Microbiota and epigenetic regulation of inflammatory mediators in type 2 diabetes and obesity. Benef. Microbes 5(1), 33–43 (2014).CAS 
    PubMed 
    Article 

    Google Scholar 
    Tamanai-Shacoori, Z. et al. Roseburia spp.: A marker of health?. Future Microbiol. 12(2), 157–170 (2017).CAS 
    PubMed 
    Article 

    Google Scholar 
    Herrmann, E. et al. RNA-based stable isotope probing suggests Allobaculum spp. as particularly active glucose assimilators in a complex murine microbiota cultured in vitro. BioMed Res. Int. 5, 1. https://doi.org/10.1155/2017/1829685 (2017).CAS 
    Article 

    Google Scholar 
    Wang, J., Wang, P., Li, D., Hu, X. & Chen, F. Beneficial effects of ginger on prevention of obesity through modulation of gut microbiota in mice. Eur. J. Nutr. 59(2), 699–718 (2020).CAS 
    PubMed 
    Article 

    Google Scholar 
    Garcia-Mazcorro, J. F., Ivanov, I., Mills, D. A. & Noratto, G. Influence of whole-wheat consumption on fecal microbial community structure of obese diabetic mice. PeerJ 4, e1702 (2016).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Wang, K. et al. Parabacteroides distasonis alleviates obesity and metabolic dysfunctions via production of succinate and secondary bile acids. Cell Rep. 26(1), 222–235 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Wu, T. R. et al. Gut commensal Parabacteroides goldsteinii plays a predominant role in the anti-obesity effects of polysaccharides isolated from Hirsutella sinensis. Gut 68(2), 248–262 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    Karl, J. P. et al. Effects of psychological, environmental and physical stressors on the gut microbiota. Front. Microbiol. 9, 2013 (2018).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    Gallè, F. et al. Exploring the association between physical activity and gut microbiota composition: a review of current evidence. Ann. Ig. Med. Prev. Comunita 31(6), 582–589 (2019).
    Google Scholar 
    Laflamme, D. R. P. C. Development and validation of a body condition score system for dogs. Canine Practice (Santa Barbara, Calif.: 1990, USA) (1997).FEDIAF. Nutritional Guidelines for Complete and Complementary Pet Food for Cats and Dogs https://fediaf.org/self-regulation/nutrition.html#guidelines (2021).Klindworth, A. et al. Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies. Nucleic Acids Res. 41(1), e1–e1 (2013).CAS 
    PubMed 
    Article 

    Google Scholar 
    Caporaso, J. G. et al. QIIME allows analysis of high-throughput community sequencing data. Nat. Methods 7(5), 335–336 (2010).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    R Core Team. R: A Language and Environment for Statistical Computing (R Foundation for Statistical Computing, 2018).
    Google Scholar 
    Lun, A. T., McCarthy, D. J. & Marioni, J. C. A step-by-step workflow for low-level analysis of single-cell RNA-seq data with bioconductor. F1000Research 5, 2122 (2016).PubMed 
    PubMed Central 

    Google Scholar 
    Gong, W., Kwak, I. Y., Pota, P., Koyano-Nakagawa, N. & Garry, D. J. DrImpute: Imputing dropout events in single cell RNA sequencing data. BMC Bioinform. 19(1), 1–10 (2018).Article 
    CAS 

    Google Scholar 
    Quast, C. et al. The SILVA ribosomal RNA gene database project: Improved data processing and web-based tools. Nucleic Acids Res. 41(D1), D590–D596 (2012).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    Finotello, F., Mastrorilli, E. & Di Camillo, B. Measuring the diversity of the human microbiota with targeted next-generation sequencing. Brief. Bioinform. 19(4), 679–692 (2018).PubMed 

    Google Scholar 
    Oksanen, J. et al. vegan: Community Ecology Package. Software http://CRAN.R-project.org/package=vegan (2012). More

  • in

    Feature selection for global tropospheric ozone prediction based on the BO-XGBoost-RFE algorithm

    Experimental dataThe dataset used in this study is the global long-term air quality indicator data of 5577 regions from 2010 to 2014 extracted by Betancourt et al.14 based on the TOAR database (https://gitlab.jsc.fz-juelich.de/esde/machine-learning/aq-bench/-/blob/master/resources/AQbench_dataset.csv)29. As shown in Fig. 3, the monitoring sites include 15 regions, including EUR (Europe), NAM (North America), and EAS (East Asia), and are mainly distributed in NAM (North America), EUR (Europe) and EAS (East Asia). The dataset mainly includes the geographical location information of the monitoring site, such as longitude and latitude, the area to which it belongs, altitude, etc., and the site environment information, such as population density, night light intensity, and vegetation coverage. Since it is difficult to directly quantify factors such as the degree of industrial activity and the degree of human activity, environmental information such as the average light intensity at night and population density are used as proxy variables for the above factors. The ozone indicator records the hourly ozone concentration from air quality observation points in various regions and aggregates the collected ozone time series in units of one year into one indicator. Using a longer aggregation period can be used to average short-term weather fluctuations. The experimental data have a total of 35 input variables, including 4 categorical attributes and 31 continuous attributes. The predictor variable is the average ozone concentration in each region from 2010 to 2014. The specific variable names and descriptions14 are shown in the supplementary materials. A total of 4/5 of the total samples were used as the training set, and 1/5 were used as the test set.Figure 3Global distribution of monitoring sites.Full size imageResults of BO-XGBoost-RFEAccording to the XGBoost-RFE algorithm for feature selection, XGBoost-RFE combined with the cross-validation method is used to calculate the selected feature set in each RFE stage for fivefold cross-validation, and the mean absolute error (MAE) is used as the evaluation criterion to finally determine the number of features with the lowest mean absolute error (MAE). At the same time, the Bayesian optimization algorithm is used to adjust the hyper-parameters of XGBoost-RFE, and then the feature subset with the lowest cross-validation mean absolute error (MAE) is obtained. The main parameters of the XGBoost model in this article include the learning_rate, n_estimators, max_depth, gamma, reg_alpha, reg_lambda, colsample_bytree, and subsample. All parameters used in the model are shown in the supplementary material. Within the given parameter range, the Bayesian optimization algorithm is used, the mean absolute error (MAE) of the XGBoost-RFE fivefold cross-validation is used as the objective function, and the number of iterations is controlled to be 100. We obtained the hyperparameter combination corresponding to the lowest MAE and the corresponding optimal feature subset. The iterative process of Bayesian optimization is shown in Fig. 4.Figure 4Iterative process of Bayesian optimization.Full size imageThe parameter range and optimized value of XGBoost-RFE are shown in Table 1. The XGBoost-RFE feature selection results under the above optimized hyperparameters are shown in Fig. 5. The number of features in the feature subset with the lowest mean absolute error is 22, and the MAE is 2.410.Table 1 Main hyper-parameter range and optimized value.Full size tableFigure 5XGBoost-RFE feature selection results: Cross-validation MAE under optimal hyperparameter combination.Full size imageAdditionally, the XGBoost-RFE feature selection model without Bayesian optimization is compared with the algorithm in this study. The default parameters of the underlying model XGBoost are set to learning_rate as 0.3, max_depth as 6, gamma as 0, colsample_bytree as 1, subsample as 1, reg_alpha as 1, and reg_lambda as 0. The comparison results are shown in Table 2. The results show that the XGBoost-RFE cross-validation MAE without parameter tuning is larger than that of the algorithm in this study, and the dimension of the feature subset obtained is also higher than that of the algorithm in this study.Table 2 Comparison of MAE and feature num before and after BO.Full size tablePrediction resultsTo test the prediction accuracy of the prediction model with the optimal subset obtained by BO-XGBoost-RFE, three indexes, MAE, RMSE and R2, are used to evaluate the prediction results, and the expressions are as follows:$$begin{array}{*{20}c} {MAE = frac{1}{n}mathop sum limits_{i = 1}^{n} left| {left( {y_{i} – widehat{{y_{i} }}} right)} right|} \ end{array}$$
    (8)
    $$begin{array}{*{20}c} {RMSE = sqrt {frac{1}{n}mathop sum limits_{i = 1}^{n} left( {y_{i} – widehat{{y_{i} }}} right)^{2} } } \ end{array}$$
    (9)
    $$begin{array}{*{20}c} {R^{2} = 1 – frac{{mathop sum nolimits_{i = 1}^{n} left( {widehat{{y_{i} }} – y_{i} } right)^{2} }}{{mathop sum nolimits_{i = 1}^{n} left( {y_{i} – overline{{y_{i} }} } right)^{2} }}} \ end{array}$$
    (10)
    n indicates the number of samples, yi is the true value, (widehat{{y_{i} }}) is the predicted value and (overline{{y_{i} }}) indicates the mean value of the predicted value.The XGBoost-RFE feature selection algorithm based on Bayesian optimization in this study is compared with feature selection using full features and features selected by the Pearson correlation coefficient, which measures the correlation between two variables. In this study, the correlation with predictor variables was selected to be less than 0.1, and the variables with correlations greater than 0.9 were deleted to avoid multicollinearity.XGBoost, random forest, support vector regression machine, and KNN algorithms were used to predict ozone concentration with full features, features selected by Pearson’s correlation coefficient, and features based on BO-XGBoost-RFE. According to the evaluation indicators described above, the comparison of the prediction performance results of the three algorithms before and after dimensionality reduction can be obtained. The MAE, RMSE and R2 results of each prediction model are shown in Table 3.Table 3 MAE, RMSE and R2 of each prediction model.Full size tableAmong the four prediction models, random forest has the lowest MAE and RMSE and the highest R2 based on three different dimensions of data and therefore has the best prediction performance. The prediction accuracy of all four prediction models based on Pearson correlation is lower than that based on BO-XGBoost-RFE, indicating that only selecting features by correlation cannot accurately extract important variables. Although the RMSE of the support vector regression model based on BO-XGBoost-RFE is slightly lower than the RMSE based on full features, the prediction accuracy of XGBoost, RF, KNN after feature selection of BO-XGBoost-RFE is higher than that based on full features and Pearson correlation. Among the four prediction models, random forest has obtained the highest prediction accuracy. The MAE based on BO-XGBoost-RFE is 5.0% and 1.4% lower than that based on the Pearson correlation coefficient and the full-feature-based model, and the RMSE is reduced by 5.1%, 1.8%, R2 improved by 4.3%, 1.4%. Additionally, the XGBoost model achieved the greatest improvement in accuracy. The MAE was reduced by 5.9% and 1.7%, the RMSE was reduced by 5.2% and 1.7%, and the R2 was improved by 4.9% and 1.4% compared with the Pearson correlation coefficient-based and full-feature-based models, respectively. This indicates that feature selection based on BO-XGBoost-RFE effectively extracts important features, improves prediction accuracy based on multiple prediction models, and has better dimensionality reduction performance.Figure 6 shows the importance of each feature obtained by using the random forest prediction model, reflecting the degree of influence of each variable on the prediction results of the global multi-year average near-ground ozone concentration. The most important variables that affect the prediction results according to the ranking of feature importance are altitude, relative altitude, and latitude, followed by night light intensity within a radius of 5 km, population density and nitrogen dioxide concentration, while the proxy variables for vegetation cover have a relatively weak effect on the prediction of ozone concentration.Figure 6Feature importance in random forest.Full size image More

  • in

    The microbiome of cryospheric ecosystems

    The datasetWe curated and explored 695 published 16S rRNA gene samples from cryospheric ecosystems (Methods section and Supplementary Table 7), including polar ice sheets, mountain glaciers and their proglacial lakes, permafrost soils and the coastal ocean under the influence of glacier runoff, and compared these to 3552 published 16S rRNA gene samples from non-cryospheric ecosystems, including temperate and tropical lakes and soils (Supplementary Table 7). This approach allowed us to identify and explore features specific to the cryospheric microbiome and compare it to other environmental microbiomes. However, we note a geographical bias towards polar regions in current publicly available repositories, and the paucity of alpine samples specifically highlights the need to further characterise these habitats given that they are among the most endangered cryospheric ecosystems globally. This bias is further compounded by the inconsistent methodologies applied across studies (e.g. primer pairs and sequencers used). To account for potential primer biases, we analysed two 16S rRNA primer pairs (Primer Pair 1, PP1: 341f-785r; Primer Pair 2, PP2: 515f-806r)12,13 commonly used in amplicon high-throughput sequencing. In total, this dataset contains 241,502,708 paired sequence reads, resulting in 530,254 and 410,931 amplicon sequence variants (ASVs) for PP1 and PP2, respectively. Moreover, all taxonomic analyses were performed at the genus level, to account for the limitations of 16s rRNA amplicon data. To gain deeper insights into the functional space of the cryospheric microbiome, we compared 34 published metagenomes from cryospheric ecosystems with 56 metagenomes from similar but non-cryospheric ecosystems (Fig. 1A). Given the difficulty of obtaining high-quality metagenomes from cryospheric ecosystems, we restricted our analyses to glacier surfaces, ice-covered lakes, and Antarctic soils. Although our analyses were limited to samples where raw sequence data are available (Methods section), the breadth of habitats covered are representative of the most abundant cryospheric ecosystems, e.g., glacier ice, cryoconites, subglacial lakes and sea ice. On the other hand, several niches such as glacier snow, glacier-fed rivers/streams, and the full-breadth of permafrost may not entirely be represented due to data unavailability. We reanalysed all metagenomes using the same bioinformatic pipeline (IMP3; see Methods section) to avoid analytical biases. Overall, the metagenomic analyses from 2,427,818,072 paired reads yielded 41,068,842 gene sequences. Thus, we here present a catalogue representing a snapshot of the functional diversity in the cryospheric microbiome, integrating across diverse habitats. This represents what we believe to be the first global overview of the functional repertoire of the Earth’s cryosphere compared to other ecosystems.Fig. 1: A unique cryospheric microbiome.A Geographic distribution of the 16 S rRNA gene samples for the two primer pairs (PP) and metagenomes for both cryospheric and non-cryospheric ecosystems, where GPS coordinates were available on NCBI. Symbol size denotes the number of samples per site (see Supplementary Table 7). B Phylogenetic tree based on abundant ASVs ( >0.5% relative abundance in at least one sample) in the PP1 dataset. The heatmap (inner rings) shows the presence (at a  > 0.5% relative abundance threshold) of ASVs in the four ecosystem types of the cryosphere (ice and snow, terrestrial, coastal ocean and freshwater). The barplot (outer ring) represents the coefficient for the SVM classifier analysis, highlighting discriminating ASVs. C Sorensen’s phylogenetic index of β-diversity (n1 = n2 = 84,461 for PP1, and n1 = n2 = 99,000 for PP2) and D β-MNTD calculated across pairs of samples in the cryospheric samples (Cryo-Cryo), pairs of cryospheric and non-cryospheric samples (Cryo-Others) and pairs of non-cryospheric (Others-Others) samples (sample sizes are listed in Supplementary Table 2). The top panel (shades of blue) is for PP1, the bottom one (shades of red) for PP2; two-sided Wilcoxon tests were performed to assess significance in panels C and D; the Holm method was used to correct for multiple testing (****: 0–0.0001). Boxplots depict the median and the 25th and 75th quartiles, whiskers extend to values within 1.5 times the interquartile range, and the remaining points are outliers. Effect sizes and exact p-values are available in Supplementary Table 2. Source data are provided as a Source Data file.Full size imageA cryospheric microbiomeGiven the communal constraints imposed by the harsh environment of cryospheric ecosystems (e.g., low temperature, oligotrophy), we expected them to harbour a specific microbiome. Accordingly, machine-learning classification (logistic regression models, Methods) based on community composition was able to differentiate between cryospheric and non-cryospheric microbiomes with high accuracy (balanced accuracy >0.96, Supplementary Table 1). Both primer pairs consistently yielded a high classification accuracy and especially a high precision. Interestingly, many of the discriminating cryospheric ASVs were spread widely across the bacterial tree of life (Fig. 1A and Supplementary Fig. 1).The notion that a part of the microbiome is specific to the cryosphere is also strongly supported by phylogenetic analyses of the 16 S rRNA gene amplicon dataset. First, we found higher pairwise phylogenetic overlap among cryospheric samples than among cryospheric/non-cryospheric or non-cryospheric samples (Sorensen’s index, Fig. 1C; Wilcoxon test, Holm adj. p  More

  • in

    The sustainability movement is 50. Why are world leaders ignoring it?

    Swedish environment minister Annika Strandhäll before the start of the Stockholm +50 Climate Summit. Few world leaders will be attending.Credit: Fredrik Persson/TT News Agency/AFP/Getty

    Sustainability is now a household term, but it wasn’t always so.Fifty years ago, the United Nations held its Conference on the Human Environment in Stockholm. This landmark event gave the concept of sustainable development its first international recognition. Sweden and the UN are marking the occasion this week with Stockholm+50, an international meeting that serves as both commemoration and call to action.The world is deep in planetary and human crises, with the UN’s Sustainable Development Goals off track and multilateral agreements on climate change and biodiversity behind schedule. Governments need to integrate sustainability into economic planning — and listen to researchers, who are ready with evidence-based arguments and tools to help them do so.Fifty years ago, the time was ripe for an environmental agenda to enter the world stage. Optimistic ideas of economic growth as a driver of progress, propelled by the Industrial Revolution, needed to accommodate concerns over damage to the natural environment. Books such as Rachel Carson’s Silent Spring (1962) — which raised awareness about harms caused by pesticides — brought scientific information about environmental risks into the mainstream.In March 1972, a team of researchers and policymakers sounded another alarm in The Limits to Growth, one of the first reports to forecast catastrophic consequences if humans kept exploiting Earth’s limited supply of natural resources. The conference in Stockholm followed a few months later, steered to success by its secretary-general, Canadian industrialist Maurice Strong. That set crucial institutions in motion, starting with the establishment of the UN Environment Programme (UNEP), based in Nairobi — the first UN body to be headquartered in a developing country. UNEP went on to facilitate a new international law — the 1987 Montreal Protocol to phase out ozone-depleting substances — and co-founded the Intergovernmental Panel on Climate Change (IPCC). It assisted in establishing the first action plans for sustainable development through landmark international agreements on biodiversity, climate and desertification.But there were mistakes and missed opportunities. The establishment of multiple agencies and policy instruments created a disjointed governance system. Newly created environment ministers wielded little power. In national budgets, environmental protection was siloed away from economic development and social concerns. For a long time, action on climate change remained unfocused. And the economic drivers of environmental change were overlooked.And so, 50 years after that momentous conference, the world remains in crisis. With impending climate and biodiversity crises, the warnings issued by visionaries now hit even closer.Stockholm+50 promises “clear and concrete recommendations and messages for action at all levels”. More than 90 ministers are expected to attend, but only 10 heads of government. That’s a missed opportunity for high-level action. World leaders are needed because their presence signals that sustainability remains at the top of their agendas.Awareness of the need to embed sustainability into policymaking has broken into the mainstream, although much of it is still talk. City governments around the world are implementing ambitious climate action plans through the C40 Cities network. Some companies, too, are adopting sustainability principles, from reporting (and reducing) their carbon footprints to ensuring that investments, as far as possible, do not harm the environment.But this urgency has not ascended to heads of state and government. With a handful of exceptions — such as Finland, Iceland, New Zealand, Scotland and Wales — most nations seem unwilling to systemically integrate their economic, environmental and social policymaking.Doing so is not only good for the environment; it is also sound economics and good for well-being. The food and energy crisis driving poverty and diminishing living standards around the world might have been triggered by the shocks of a pandemic and war on Ukraine — but it is driven just as much by the depletion of natural resources.Ahead of the 1972 conference, 2,200 environmental scientists signed a letter — called the Menton Message — to then UN secretary-general U Thant. The signatories had a sense that the world was moving towards multiple crises. They urged “massive research into the problems that threaten the survival of mankind”, such as hunger, wars, environmental degradation and natural-resource depletion. The UN system went on to play a big part in building the body of knowledge that has shown why sustainability is necessary, and in creating the policy architecture to make it happen. But to do the Stockholm vision justice, there must be bolder action from heads of government and from the UN system. The planned creation of a board of science advisers to UN secretary-general António Guterres needs to be accelerated. Once established, the board must find a way to bring joined-up action on sustainability closer to world leaders.Researchers can now join a successor to the Menton Message that has been organized by the International Science Council, the global science network Future Earth and the Stockholm Environment Institute. In an open letter addressed to world citizens, the authors write: “After 50 years, pro-environmental action seems like one step forward and two back. The world produces more food than needed, yet many people still go hungry. We continue to subsidize and invest in fossil fuels, even though renewable energy is increasingly cost-effective. We extract resources where the price is lowest, often in direct disregard of local rights and values.”World leaders must listen to the research community, and accept the evidence and narrative offered to help them to navigate meaningful change. Environmental sustainability does not impede prosperity and well-being — in fact, it is vital to them. People in power need to sit up and take notice. More

  • in

    Success of post-fire plant recovery strategies varies with shifting fire seasonality

    Canadell, J. G. et al. Multi-decadal increase of forest burned area in Australia is linked to climate change. Nat. Commun. 12, 6921 (2021).CAS 
    Article 

    Google Scholar 
    Jolly, W. M. et al. Climate-induced variations in global wildfire danger from 1979 to 2013. Nat. Commun. 6, 7537 (2015).CAS 
    Article 

    Google Scholar 
    Jain, P., Wang, X. & Flannigan, M. D. Trend analysis of fire season length and extreme fire weather in North America between 1979 and 2015. Int. J. Wildland Fire 26, 1009–1020 (2017).Article 

    Google Scholar 
    Wotton, B. M. & Flannigan, M. D. Length of the fire season in a changing climate. For. Chronicle 69, 187–192 (1993).
    Google Scholar 
    Collins, L. et al. The 2019/2020 mega-fires exposed Australian ecosystems to an unprecedented extent of high-severity fire. Environ. Res. Lett. 16, 044029 (2021).Article 

    Google Scholar 
    Higuera, P. E. & Abatzoglou, J. T. Record‐setting climate enabled the extraordinary 2020 fire season in the western United States. Glob. Change Biol. 27, 1–2 (2021).Article 

    Google Scholar 
    Nolan, R. H. et al. Limits to post-fire vegetation recovery under climate change. Plant Cell Environ. 44, 3471–3489 (2021).CAS 
    Article 

    Google Scholar 
    Abram, N. J. et al. Connections of climate change and variability to large and extreme forest fires in southeast Australia. Commun. Earth Environ. 2, 8 (2021).Article 

    Google Scholar 
    Dickman, C. R. Ecological consequences of Australia’s “Black Summer” bushfires: managing for recovery. Int. Environ. Assess. Manag. 17, 1162–1167 (2021).Article 

    Google Scholar 
    Swain, D. L. A shorter, sharper rainy season amplifies California wildfire risk. Geophys. Res. Lett. 48, e2021GL092843 (2021).
    Google Scholar 
    Keeley, J. E. Fire intensity, fire severity and burn severity: a brief review and suggested usage. Int. J. Wildland Fire 18, 116–126 (2009).Article 

    Google Scholar 
    He, T., Pausas, J. G., Belcher, C. M., Schwilk, D. W. & Lamont, B. B. Fire-adapted traits of Pinus arose in the fiery Cretaceous. New Phytol. 194, 751–759 (2012).Article 

    Google Scholar 
    Bradstock, R. A. A biogeographic model of fire regimes in Australia: current and future implications. Glob. Ecol. Biogeogr. 19, 145–158 (2010).Article 

    Google Scholar 
    Bowman, D. M., Murphy, B. P., Neyland, D. L., Williamson, G. J. & Prior, L. D. Abrupt fire regime change may cause landscape-wide loss of mature obligate seeder forests. Glob. Change Biol. 20, 1008–1015 (2014).Article 

    Google Scholar 
    Keeley, J. E., Pausas, J. G., Rundel, P. W., Bond, W. J. & Bradstock, R. A. Fire as an evolutionary pressure shaping plant traits. Trends Plant Sci. 16, 406–411 (2011).CAS 
    Article 

    Google Scholar 
    Barrett, K. et al. Postfire recruitment failure in Scots pine forests of southern Siberia. Remote Sens. Environ. 237, 111539 (2020).Article 

    Google Scholar 
    Miller, R. G., Fontaine, J. B., Merritt, D. J., Miller, B. P. & Enright, N. J. Experimental seed sowing reveals seedling recruitment vulnerability to unseasonal fire. Ecol. Appl. 31, e02411 (2021).
    Google Scholar 
    Prior, L. D., Williamson, G. J. & Bowman, D. M. Impact of high-severity fire in a Tasmanian dry eucalypt forest. Austral. J. Bot. 64, 193–205 (2016).Article 

    Google Scholar 
    Brewer, J. S. Long-term population changes of a fire-adapted plant subjected to different fire seasons. Nat. Areas J. 26, 267–273 (2006).Article 

    Google Scholar 
    Keith, D. A., Holman, L., Rodoreda, S., Lemmon, J. & Bedward, M. Plant functional types can predict decade‐scale changes in fire‐prone vegetation. J. Ecol. 95, 1324–1337 (2007).Article 

    Google Scholar 
    Savage, M., Mast, J. N. & Feddema, J. J. Double whammy: high-severity fire and drought in ponderosa pine forests of the Southwest. Can. J. For. Res. 43, 570–583 (2013).Article 

    Google Scholar 
    Miller, R. G. et al. Mechanisms of fire seasonality effects on plant populations. Trends Ecol. Evol. 34, 1104–1117 (2019).Article 

    Google Scholar 
    Tangney, R., Merritt, D. J., Fontaine, J. B. & Miller, B. P. Seed moisture content as a primary trait regulating the lethal temperature thresholds of seeds. J. Ecol. 107, 1093–1105 (2019).Article 

    Google Scholar 
    Tangney, R. et al. Seed dormancy interacts with fire seasonality mechanisms. Trends Ecol. Evol. 35, 1057–1059 (2020).Article 

    Google Scholar 
    Bowman, D. M. et al. The human dimension of fire regimes on Earth. J. Biogeogr. 38, 2223–2236 (2011).Article 

    Google Scholar 
    Knapp, E. E., Estes, B. L. & Skinner, C. N. Ecological effects of prescribed fire season: a literature review and synthesis for managers. Gen. Tech. Rep. https://doi.org/10.2737/PSW-GTR-224 (2009).Miller, R. G. et al. Fire seasonality mechanisms are fundamental for understanding broader fire regime effects. Trends Ecol. Evol. 35, 869–871 (2020).Article 

    Google Scholar 
    Keeley, J. E. & Syphard, A. D. Twenty-first century California, USA, wildfires: fuel-dominated vs. wind-dominated fires. Fire Ecol. 15, 24 (2019).Article 

    Google Scholar 
    Lamont, B. B., Enright, N. J. & He, T. Fitness and evolution of resprouters in relation to fire. Plant Ecol. 212, 1945–1957 (2011).Article 

    Google Scholar 
    Pausas, J. G. & Bradstock, R. A. Fire persistence traits of plants along a productivity and disturbance gradient in mediterranean shrublands of south‐east Australia. Glob. Ecol. Biogeogr. 16, 330–340 (2007).Article 

    Google Scholar 
    Pausas, J. G. & Keeley, J. E. Evolutionary ecology of resprouting and seeding in fire-prone ecosystems. New Phytol. 204, 55–65 (2014).Article 

    Google Scholar 
    Fairman, T. A., Bennett, L. T. & Nitschke, C. R. Short-interval wildfires increase likelihood of resprouting failure in fire-tolerant trees. J. Environ. Manag. 231, 59–65 (2019).Article 

    Google Scholar 
    Pyke, G. H. Fire-stimulated flowering: a review and look to the future. Critic. Rev. Plant Sci. 36, 179–189 (2017).Article 

    Google Scholar 
    Zirondi, H. L., Ooi, M. K. J. & Fidelis, A. Fire-triggered flowering is the dominant post-fire strategy in a tropical savanna. J. Veg. Sci. 32, e12995 (2021).Article 

    Google Scholar 
    Howe, H. F. Response of Zizia aurea to seasonal mowing and fire in a restored Prairie. Am. Midl. Nat. 141, 373–380 (1999).Article 

    Google Scholar 
    Thompson, K. Seeds and seed banks. New Phytol. 106, 23–34 (1987).Article 

    Google Scholar 
    Baskin, C. C. & Baskin, J. M. Seeds: Ecology, Biogeography, and Evolution of Dormancy and Germination 2nd edn (Academic Press, 2001).Alvarado, V. & Bradford, K. J. A hydrothermal time model explains the cardinal temperatures for seed germination. Plant Cell Environ. 25, 1061–1069 (2002).Article 

    Google Scholar 
    Mackenzie, B. D. E., Auld, T. D., Keith, D. A., Hui, F. K. C. & Ooi, M. K. J. The effect of seasonal ambient temperatures on fire-stimulated germination of species with physiological dormancy: a case study using boronia (Rutaceae). PLoS One 11, e0156142 (2016).Article 
    CAS 

    Google Scholar 
    Ooi, M. K. J. Delayed emergence and post-fire recruitment success: effects of seasonal germination, fire season and dormancy type. Austral. J. Bot. 58, 248–256 (2010).Article 

    Google Scholar 
    Bond, W. Fire survival of Cape Proteaceae-influence of fire season and seed predators. Vegetatio 56, 65–74 (1984).Article 

    Google Scholar 
    Keith, D. A., Dunker, B. & Driscoll, D. A. Dispersal: the eighth fire seasonality effect on plants. Trends Ecol. Evol. 35, 305–307 (2020).Article 

    Google Scholar 
    Paroissien, R. & Ooi, M. K. J. Effects of fire season on the reproductive success of the post-fire flowerer Doryanthes excelsa. Environ. Exp. Bot. 192, 104634 (2021).Article 

    Google Scholar 
    Furlaud, J. M., Prior, L. D., Williamson, G. J. & Bowman, D. M. J. S. Bioclimatic drivers of fire severity across the Australian geographical range of giant Eucalyptus forests. J. Ecol. 109, 2514–2536 (2021).Article 

    Google Scholar 
    Thomsen, A. M. & Ooi, M. K. J. Shifting season of fire and its interaction with fire severity: Impacts on reproductive effort in resprouting plants. Ecol. Evol. 12, e8717 (2022).Article 

    Google Scholar 
    Fill, J. M. & Crandall, R. M. Stronger evidence needed for global fire season effects. Trends Ecol. Evol. 35, 867–868 (2020).Article 

    Google Scholar 
    Jump, A. S. & Peñuelas, J. Running to stand still: adaptation and the response of plants to rapid climate change. Ecol. Lett. 8, 1010–1020 (2005).Article 

    Google Scholar 
    Inouye, D. W. Climate change and phenology. Wiley Interdiscip. Rev. Clim. Change n/a, e764 (2022).
    Google Scholar 
    Enright, N. J., Marsula, R., Lamont, B. B. & Wissel, C. The ecological significance of canopy seed storage in fire-prone environments: a model for non-sprouting shrubs. J. Ecol. 86, 946–959 (1998).Article 

    Google Scholar 
    Setterfield, S. A. The impact of experimental fire regimes on seed production in two tropical eucalypt species in northern Australia. Austral. J. Ecol. 22, 279–287 (1997).Article 

    Google Scholar 
    Collette, J. C. & Ooi, M. K. J. Evidence for physiological seed dormancy cycling in the woody shrub Asterolasia buxifolia and its ecological significance in fire-prone systems. Plant Biol. 22, 745–749 (2020).CAS 
    Article 

    Google Scholar 
    Setterfield, S. A. Seedling establishment in an Australian tropical savanna: effects of seed supply, soil disturbance and fire. J. Appl. Ecol. 39, 949–959 (2002).Article 

    Google Scholar 
    Russell-Smith, J. & Edwards, A. C. Seasonality and fire severity in savanna landscapes of monsoonal northern Australia. Int. J. Wildland Fire 15, 541–550 (2006).Article 

    Google Scholar 
    Whitehead, P. J., Purdon, P., Russell-Smith, J., Cooke, P. M. & Sutton, S. The management of climate change through prescribed Savanna burning: Emerging contributions of indigenous people in Northern Australia. Public Adm. Dev. 28, 374–385 (2008).Article 

    Google Scholar 
    Prior, L. D., Williams, R. J. & Bowman, D. M. Experimental evidence that fire causes a tree recruitment bottleneck in an Australian tropical savanna. J. Tropical Ecol. 26, 595–603 (2010).Abatzoglou, J. T., Williams, A. P. & Barbero, R. Global emergence of anthropogenic climate change in fire weather indices. Geophys. Res. Lett. 46, 326–336 (2019).Article 

    Google Scholar 
    Ferreira, L. N., Vega-Oliveros, D. A., Zhao, L., Cardoso, M. F. & Macau, E. E. N. Global fire season severity analysis and forecasting. Comput. Geosci. 134, 104339 (2020).Article 

    Google Scholar 
    Flannigan, M. et al. Global wildland fire season severity in the 21st century. For. Ecol. Manag. 294, 54–61 (2013).Article 

    Google Scholar 
    Ansley, R. J. & Castellano, M. J. Prickly pear cactus responses to summer and winter fires. Rangel. Ecol. Manag. 60, 244–252 (2007).Article 

    Google Scholar 
    Ansley, R. J., Kramp, B. A. & Jones, D. L. Honey mesquite (Prosopis glandulosa) seedling responses to seasonal timing of fire and fireline intensity. Rangel. Ecol. Manag. 68, 194–203 (2015).Article 

    Google Scholar 
    Armstrong, G. & Legge, S. The post-fire response of an obligate seeding Triodia species (Poaceae) in the fire-prone Kimberley, north-west Australia. Int. J. Wildland Fire 20, 974–981 (2012).Article 

    Google Scholar 
    Bellows, R. S., Thomson, A. C., Helmstedt, K. J., York, R. A. & Potts, M. D. Damage and mortality patterns in young mixed conifer plantations following prescribed fires in the Sierra Nevada, California. For. Ecol. Manag. 376, 193–204 (2016).Article 

    Google Scholar 
    Beyers, J. L. & Wakeman, C. D. Season of burn effects in southern California chaparral. In Second interface between ecology and land development in California 45–55 (Occidental College, CA, 2000).Bowen, B. J. & Pate, J. S. Effect of season of burn on shoot recovery and post‐fire flowering performance in the resprouter Stirlingia latifolia R. Br.(Proteaceae). Austral Ecol. 29, 145–155 (2004).Article 

    Google Scholar 
    Casals, P., Valor, T., Rios, A. & Shipley, B. Leaf and bark functional traits predict resprouting strategies of understory woody species after prescribed fires. For. Ecol. Manag. 429, 158–174 (2018).Article 

    Google Scholar 
    Céspedes, B., Torres, I., Luna, B., Pérez, B. & Moreno, J. M. Soil seed bank, fire season, and temporal patterns of germination in a seeder-dominated Mediterranean shrubland. Plant Ecol. 213, 383–393 (2012).Article 

    Google Scholar 
    Clabo, D. C. & Clatterbuck, W. K. Shortleaf pine (Pinus echinata, Pinaceae) seedling sprouting responses: Clipping and burning effects at various seedling ages and seasons. J. Torrey Bot. Soc. 146, 96–110 (2019).Article 

    Google Scholar 
    Drewa, P. B. Effects of fire season and intensity on Prosopis glandulosa Torr. var. glandulosa. Int. J. Wildland Fire 12, 147–157 (2003).Article 

    Google Scholar 
    Drewa, P. B., Platt, W. J. & Moser, E. B. Fire effects on resprouting of shrubs in headwaters of southeastern longleaf pine savannas. Ecology 83, 755–767 (2002).Article 

    Google Scholar 
    Drewa, P. B., Thaxton, J. M. & Platt, W. J. Responses of root‐crown bearing shrubs to differences in fire regimes in Pinus palustris (longleaf pine) savannas: exploring old‐growth questions in second‐growth systems. Appl. Veg. Sci. 9, 27–36 (2006).
    Google Scholar 
    Ellsworth, L. M. & Kauffman, J. B. Seedbank responses to spring and fall prescribed fire in mountain big sagebrush ecosystems of differing ecological condition at Lava Beds National Monument, California. J. Arid Environ. 96, 1–8 (2013).Article 

    Google Scholar 
    Fairfax, R. et al. Effects of multiple fires on tree invasion in montane grasslands. Landsc. Ecol. 24, 1363–1373 (2009).Article 

    Google Scholar 
    Fill, J. M., Welch, S. M., Waldron, J. L. & Mousseau, T. A. The reproductive response of an endemic bunchgrass indicates historical timing of a keystone process. Ecosphere 3, 1–12 (2012).Article 

    Google Scholar 
    Grant, C. Post-burn vegetation development of rehabilitated bauxite mines in Western Australia. For. Ecol. Manag. 186, 147–157 (2003).Article 

    Google Scholar 
    Hajny, K. M., Hartnett, D. C. & Wilson, G. W. Rhus glabra response to season and intensity of fire in tallgrass prairie. Int. J. Wildland Fire 20, 709–720 (2011).Article 

    Google Scholar 
    Holmes, P. A comparison of the impacts of winter versus summer burning of slash fuel in alien-invaded fynbos areas in the Western Cape. Southern African For. J. 192, 41–50 (2001).Article 

    Google Scholar 
    Jasinge, N., Huynh, T. & Lawrie, A. Consequences of season of prescribed burning on two spring-flowering terrestrial orchids and their endophytic fungi. Austr. J. Bot. 66, 298–312 (2018).Article 

    Google Scholar 
    Jasinge, N., Huynh, T. & Lawrie, A. Changes in orchid populations and endophytic fungi with rainfall and prescribed burning in Pterostylis revoluta in Victoria, Australia. Ann. Bot. 121, 321–334 (2018).CAS 
    Article 

    Google Scholar 
    Kauffman, J. & Martin, R. Sprouting shrub response to different seasons and fuel consumption levels of prescribed fire in Sierra Nevada mixed conifer ecosystems. For. Sci. 36, 748–764 (1990).
    Google Scholar 
    Keyser, T. L., Greenberg, C. H. & McNab, W. H. Season of burn effects on vegetation structure and composition in oak-dominated Appalachian hardwood forests. For. Ecol. Manag. 433, 441–452 (2019).Article 

    Google Scholar 
    Knox, K. & Clarke, P. J. Fire season and intensity affect shrub recruitment in temperate sclerophyllous woodlands. Oecologia 149, 730–739 (2006).CAS 
    Article 

    Google Scholar 
    Lamont, B. B. & Downes, K. S. Fire-stimulated flowering among resprouters and geophytes in Australia and South Africa. Plant Ecol. 212, 2111–2125 (2011).Article 

    Google Scholar 
    Lesica, P. & Martin, B. Effects of prescribed fire and season of burn on recruitment of the invasive exotic plant, Potentilla recta, in a semiarid grassland. Restoration Ecol. 11, 516–523 (2003).Article 

    Google Scholar 
    Moreno, J. M. et al. Rainfall patterns after fire differentially affect the recruitment of three Mediterranean shrubs. Biogeosciences 8, 3721–3732 (2011).Article 

    Google Scholar 
    Mulligan, M. K. & Kirkman, L. K. Burning influences on wiregrass (Aristida beyrichiana) restoration plantings: natural seedling recruitment and survival. Restor. Ecol. 10, 334–339 (2002).Article 

    Google Scholar 
    Nield, A. P., Enright, N. J. & Ladd, P. G. Fire-stimulated reproduction in the resprouting, non-serotinous conifer Podocarpus drouynianus (Podocarpaceae): the impact of a changing fire regime. Popul. Ecol. 58, 179–187 (2016).Article 

    Google Scholar 
    Norden, A. H. & Kirkman, L. K. Persistence and prolonged winter dormancy of the federally endangered Schwalbea Americana L.(Scrophulariaceae) following experimental management techniques. Nat. Areas J. 24, 129–134 (2004).
    Google Scholar 
    Olson, M. S. & Platt, W. J. Effects of habitat and growing season fires on resprouting of shrubs in longleaf pine savannas. Vegetatio 119, 101–118 (1995).Article 

    Google Scholar 
    Ooi, M. K. The importance of fire season when managing threatened plant species: a long-term case-study of a rare Leucopogon species (Ericaceae). J. Environ. Manag. 236, 17–24 (2019).Article 

    Google Scholar 
    Pavlovic, N. B., Leicht-Young, S. A. & Grundel, R. Short-term effects of burn season on flowering phenology of savanna plants. Plant Ecology 212, 611–625 (2011).Article 

    Google Scholar 
    Payton, I. J. & Pearce, H. G. Fire-Induced Changes to the Vegetation of Tall-Tussock (Chionochloa rigida) Grassland Ecosystems. (Department of Conservation Wellington, New Zealand, 2009).Peguero, G. & Espelta, J. M. Disturbance intensity and seasonality affect the resprouting ability of the neotropical dry-forest tree Acacia pennatula: do resources stored below-ground matter? J. Tropical Ecol. 28, 539–546 (2011).Risberg, L. & Granström, A. Exploiting a window in time. Fate of recruiting populations of two rare fire-dependent Geranium species after forest fire. Plant Ecol. 215, 613–624 (2014).Article 

    Google Scholar 
    Rodríguez-Trejo, D. A., Castro-Solis, U. B., Zepeda-Bautista, M. & Carr, R. J. First year survival of Pinus hartwegii following prescribed burns at different intensities and different seasons in central Mexico. Int. J. Wildland Fire 16, 54–62 (2007).Article 

    Google Scholar 
    Russell, M., Vermeire, L., Ganguli, A. & Hendrickson, J. Fire return interval and season of fire alter bud banks. Rangel. Ecol. Manag.72, 542–550 (2019).Article 

    Google Scholar 
    Russell-Smith, J., Whitehead, P. J., Cook, G. D. & Hoare, J. L. Response of Eucalyptus‐dominated savanna to frequent fires: lessons from Munmarlary, 1973–1996. Ecol. Monogr. 73, 349–375 (2003).Article 

    Google Scholar 
    Schmidt, I. B., Sampaio, A. B. & Borghetti, F. Effects of the season on sexual reproduction and population structure of Heteropterys pteropetala (Adr. Juss.), Malpiguiaceae, in areas of Cerrado sensu stricto submitted to biennial fires. Acta Bot. Brasilica 19, 927–934 (2005).Article 

    Google Scholar 
    Shepherd, B. J., Miller, D. L. & Thetford, M. Fire season effects on flowering characteristics and germination of longleaf pine (Pinus palustris) savanna grasses. Restor. Ecol. 20, 268–276 (2012).Article 

    Google Scholar 
    Spier, L. P. & Snyder, J. R. Effects of wet-and dry-season fires on Jacquemontia curtisii, a south Florida pine forest endemic. Nat. Areas J. 18, 350–357 (1998).
    Google Scholar 
    Tsafrir, A. et al. Fire season modifies the perennial plant community composition through a differential effect on obligate seeders in eastern Mediterranean woodlands. Appl. Veg. Sci. 22, 115–126 (2019).Article 

    Google Scholar 
    Vander Yacht, A. L. et al. Vegetation response to canopy disturbance and season of burn during oak woodland and savanna restoration in Tennessee. For. Ecol. Manag. 390, 187–202 (2017).Article 

    Google Scholar 
    Vidaller, C., Dutoit, T., Ramone, H. & Bischoff, A. Fire increases the reproduction of the dominant grass Brachypodium retusum and Mediterranean steppe diversity in a combined burning and grazing experiment. Appl. Veg. Sci. 22, 127–137 (2019).Article 

    Google Scholar 
    Williams, P. R., Congdon, R. A., Grice, A. C. & Clarke, P. J. Soil temperature and depth of legume germination during early and late dry season fires in a tropical eucalypt savanna of north‐eastern Australia. Austral Ecol. 29, 258–263 (2004).Article 

    Google Scholar 
    Williams, P. R., Congdon, R. A., Grice, A. C. & Clarke, P. J. Germinable soil seed banks in a tropical savanna: seasonal dynamics and effects of fire. Austral Ecol. 30, 79–90 (2005).Article 

    Google Scholar 
    Zhao, H. et al. Ecophysiological influences of prescribed burning on wetland plants: a case study in Sanjiang Plain wetlands, northeast China. Fresenius Environ. Bull 20, 2932–2938 (2011).CAS 

    Google Scholar 
    Pick, J. L., Nakagawa, S. & Noble, D. W. Reproducible, flexible and high‐throughput data extraction from primary literature: The metaDigitise r package. Methods in Ecol. Evol. 10, 426–431 (2019).Article 

    Google Scholar 
    Team, R. C. R: A Language and Environment for Statistical Computing. https://www.R-project.org/ (2020).Higgins, J. P. et al. Cochrane Handbook for Systematic Reviews of Interventions. (John Wiley & Sons, 2019).Lüdecke, D., Lüdecke, M. D. & David, B. W. Package ‘esc’. https://strengejacke.github.io/esc (2017).Schwarzer, G. meta: An R package for meta-analysis. R news 7, 40–45 (2007).
    Google Scholar 
    Borenstein, M., Hedges, L. V., Higgins, J. P. & Rothstein, H. R. A basic introduction to fixed‐effect and random‐effects models for meta‐analysis. Res. Synth. Methods 1, 97–111 (2010).Article 

    Google Scholar 
    Harrer, M., Cuijpers, P., Furukawa, T. A. & Ebert, D. D. Doing Meta-Analysis with R: a Hands-on Guide. (Chapman and Hall, 2019).Wilke, C. O., Wickham, H. & Wilke, M. C. O. Package ‘cowplot’. Streamlined Plot Theme and Plot Annotations for ‘ggplot2 (Cowplot, 2019).Fill, J. M., Davis, C. N. & Crandall, R. M. Climate change lengthens southeastern USA lightning‐ignited fire seasons. Glob. Change Biol. 25, 3562–3569 (2019).Article 

    Google Scholar 
    Halofsky, J. E., Peterson, D. L. & Harvey, B. J. Changing wildfire, changing forests: the effects of climate change on fire regimes and vegetation in the Pacific Northwest, USA. Fire Ecol. 16, 4 (2020).Article 

    Google Scholar 
    Kraaij, T., Cowling, R. M., van Wilgen, B. W., Rikhotso, D. R. & Difford, M. Vegetation responses to season of fire in an aseasonal, fire-prone fynbos shrubland. PeerJ 5, e3591 (2017).Article 

    Google Scholar 
    Peel, M. C., Finlayson, B. L. & McMahon, T. A. Updated world map of the Köppen-Geiger climate classification. Hydrol. Earth Syst. Sci. 11, 1633–1644 (2007).Article 

    Google Scholar 
    Murphy, B. P. et al. Fire regimes of Australia: a pyrogeographic model system. J. Biogeogr. 40, 1048–1058 (2013).Article 

    Google Scholar 
    McColl-Gausden, S. C., Bennett, L. T., Duff, T. J., Cawson, J. G. & Penman, T. D. Climatic and edaphic gradients predict variation in wildland fuel hazard in south-eastern Australia. Ecography 43, 443–455 (2020).Article 

    Google Scholar 
    Pausas, J. G. & Keeley, J. E. Evolutionary ecology of resprouting and seeding in fire‐prone ecosystems. New Phytol. 204, 55–65 (2014).Article 

    Google Scholar 
    Lamont, B. B., Maitre, D. C. L., Cowling, R. M. & Enright, N. J. Canopy seed storage in woody plants. Bot. Rev. 57, 277–317 (1991).Article 

    Google Scholar 
    Tangney, R. et al. Data supporting: Success of post-fire plant recovery strategies varies with shifting fire seasonality. Zenodo https://doi.org/10.5061/dryad.7sqv9s4t5 (2022).Rothstein, H. R., Sutton, A. J. & Borenstein, M. Publication Bias in Meta-Analysis: Prevention, Assessment and Adjustments (John Wiley & Sons, 2006).Head, M. L., Holman, L., Lanfear, R., Kahn, A. T. & Jennions, M. D. The extent and consequences of P-hacking in science. PLoS Biol. 13, e1002106 (2015).Article 
    CAS 

    Google Scholar 
    Egger, M., Smith, G. D., Schneider, M. & Minder, C. Bias in meta-analysis detected by a simple, graphical test. BMJ 315, 629–634 (1997).CAS 
    Article 

    Google Scholar 
    Simonsohn, U., Nelson, L. D. & Simmons, J. P. P-curve: a key to the file-drawer. J. Exp. Psychol.Gen. 143, 534 (2014).Article 

    Google Scholar  More

  • in

    Influence of spatial characteristics of green spaces on microclimate in Suzhou Industrial Park of China

    In this study, the five main characteristics of green spaces that were measured were area, perimeter, perimeter-area ratio, leaf area index, and canopy density. The structure of parameter between them is shown in Table 3.Table 3 Parameter structure of the cooling and humidification effect based on the spatial characteristics of green spaces.Full size tableCorrelation between various spatial characteristics and cooling and humidifying intensity in green spacesSmall-size green spacesFigures 4 and 6 shows the results of linear regressions between spatial characteristics and the cooling effect in small-size green spaces. There were relatively weak correlations between area, perimeter, perimeter-area ratio, leaf area index and cooling intensity, and a strong correlation between canopy density and cooling intensity. Small-size green space has the weakest positive correlation between perimeter-area ratio and cooling intensity (R2 = 0.11), and its canopy density and cooling intensity have the strongest positive correlation (R2 = 0.64). Meanwhile, small-size green space has weakest negative correlation between perimeter and humidifying intensity (R2 = 0.17), and its leaf area index and humidifying intensity have significant positive correlation (R2 = 0.42). Figures 4a and 5a show that for every 1 ha increase in area of small-size green spaces, the cooling intensity increased by 1.026 °C, and the humidifying intensity decreased by 1.56%. Figures 4b and 5b show that for every 100 m increase in perimeter, the cooling intensity decreases by 1.06 °C, and the humidifying intensity decreased by 1.19%. Figures 4c and 5c show that for every 0.01 increase in the perimeter-area ratio, the cooling intensity increases by 1.12 °C, and the humidifying intensity increased by 1.46%. Figures 4d and 5d show that for every 0.1 increase in the leaf area index, the cooling intensity increases by 1.11 °C, and the humidifying intensity increased by 1.12%. Figures 4e and 5e show that each 0.01 increase in the canopy density, the cooling intensity increases by 1.60 °C, and each 0.1 increase in canopy density, the humidifying intensity increased by 1.15% (Fig. 6).
    Figure 4Linear regressions between spatial characteristics and cooling intensity of small-size green spaces.Full size imageFigure 5Linear regressions of spatial characteristics and humidifying intensity of small-size green spaces.Full size imageFigure 6The correlation between the spatial characteristics of small-size green spaces and the intensity of cooling and humidifying (GA means green area; GP means green perimeter; GPAR means green perimeter-area ratio; LAI means leaf area index; CD means canopy density).Full size imageMedium-size green spacesFigures 7 and 9 shows the linear regressions between spatial characteristics and cooling intensity in medium-size green spaces. There was an extremely significant positive correlation between area and cooling intensity, an insignificant positive correlation between the leaf area index and cooling intensity, and a relatively weak negative correlation between the other three characteristics and cooling intensity. Medium-size green space has the weakest negative correlation between canopy density and cooling intensity (R2 = 0.12), and its green area and cooling intensity have the strongest positive correlation (R2 = 0.83). Meanwhile, medium-size green space has weakest negative correlation between perimeter-area ratio and humidifying intensity (R2 = 0.41), and its area and humidifying intensity have most significant positive correlation (R2 = 0.81). Figures 7a and 8a show that for every 1 ha increase in area of medium-size green spaces, the cooling intensity increased by 1.19 °C, and the humidifying intensity increased by 1.24%. Figures 7b and 8b show that for every 100 m increase in perimeter, the cooling intensity decreases by 1.02 °C, and the humidifying intensity increased by 1.17%. Figures 7c and 8c show that for every 0.01 increase in the perimeter-area ratio, the cooling intensity decreases by 1.29 °C, and the humidifying intensity decreased by 2.40%. Figures 7d and 8d show that for every 0.1 increase in the leaf area index, the cooling intensity increases by 1.37 °C, and the humidifying intensity decreased by 1.92%. Figures 7e and 8e show that each 0.01 increase in the canopy density, increases the cooling intensity decreases by 1.23 °C, and the humidifying intensity decreased by 6.48% (Fig. 9).Figure 7Linear regressions between spatial characteristics and cooling intensity of medium-size green spaces.Full size imageFigure 8Linear regressions of spatial characteristics and humidifying intensity of medium-size green spaces.Full size imageFigure 9The correlation between the spatial characteristics of medium-size green spaces and the intensity of cooling and humidifying (GA means green area; GP means green perimeter; GPAR means green perimeter-area ratio; LAI means leaf area index; CD means canopy density).Full size imageLarge-size green spacesFigures 10 and 12 shows the linear regressions between spatial characteristics and cooling intensity in large-size green spaces. There was an insignificant correlation between area and cooling intensity, a weak correlation between canopy density and cooling intensity, and a significant correlation between perimeter, perimeter-area ratio and the leaf area index and cooling intensity. Medium-size green space has the weakest negative correlation between green area and cooling intensity (R2 = 0.35), and its leaf area index and cooling intensity have the strongest positive correlation (R2 = 0.92). Meanwhile, medium-size green space has weakest negative correlation between perimeter-area ratio and humidifying intensity (R2 = 0.11), and its leaf area index and humidifying intensity have most significant positive correlation (R2 = 0.39). Figures 10a and 11a show that for every 1 ha increase in area of large-size green spaces, the cooling intensity decreased by 1.02 °C, and the humidifying intensity decreased by 1.22%. Figures 10b and 11b show that for every 100 m increase in perimeter, the cooling intensity decreases by 1.05 °C, and the humidifying intensity decreased by 1.34%. Figures 10c and 11c show that for every 0.005 increase in the perimeter-area ratio, the cooling intensity decreases by 1.43 °C, and each 0.01 increase in perimeter-area ratio, the humidifying intensity decreased by 1.27%. Figures 10d and 11d show that for every 0.1 increase in the leaf area index, the cooling intensity increases by 2.41 °C, and the humidifying intensity increased by 1.37%. Figures 10e and 11e show that each 0.1 increase in the canopy density, the cooling intensity increased by 3.69 °C, and the humidifying intensity decreased by 2.84% (Fig. 12).Figure 10Linear regressions of spatial characteristics and cooling intensity of large-size green spaces.Full size imageFigure 11Linear regressions of spatial characteristics and humidifying intensity of large-size green spaces.Full size imageFigure 12The correlation between the spatial characteristics of large-size green spaces and the intensity of cooling and humidifying (GA means green area; GP means green perimeter; GPAR means green perimeter-area ratio; LAI means leaf area index; CD means canopy density).Full size imageQuantitative analysis of the microclimatic effects of different types of green spacesQuantitative analysis of the effects of different types of green space on cooling intensityFigure 13 shows the linear regressions between the different types of green spaces and cooling intensity. There were negative correlations between green spaces a short, medium, and long distance from a water body and cooling intensity in small-size green spaces, medium-size green spaces and large-size green spaces. The negative correlation between the distance to a water body and cooling intensity in medium-size green spaces was most significant (R2 = 0.985). The greater the distance to a water body, the lower the cooling intensity. For medium-size green spaces, for every 1/4 increase in the distance ratio, the cooling intensity decreased by 0.81 °C. For small-size green spaces, for every 1/4 increase in the distance ratio, the cooling intensity decreased by 1.04 °C. For large-size green spaces, for every 1/4 increase in the distance ratio, the cooling intensity decreased by 1.36 °C. For small-, medium-, and large-size green spaces, there was a positive correlation between canopy density and cooling intensity. There was a most significant positive correlation between canopy density and cooling intensity in large-size green spaces (R2 = 0.941). The greater the canopy density, the greater the cooling intensity. For large green spaces, for every 0.5 increase in canopy density, the cooling intensity increased by 0.16 °C. For small-size green spaces, for every 0.5 increase in canopy density, the cooling effect increased by 0.15 °C. For medium-size green spaces, for every 0.5 increase in canopy density, the cooling intensity increased by 0.16 °C.Figure 13Linear regressions between the distance from different types of green spaces to water areas, canopy density and cooling intensity.Full size imageQuantitative analysis of the effects of different types of green space on humidifying intensityFigure 14 shows the linear regression between the distance of a green space from a water body, canopy density and humidifying intensity. There was a negative correlation between the distance to a water body and humidifying intensity in small, medium, and large green spaces. The negative correlation between the distance to a water body and humidifying intensity in small green spaces was most significant (R2 = 0.996). The longer the distance, the lower the humidifying intensity. For small green spaces, for every 1/4 in-crease in the distance ratio, the humidifying intensity decreased by 4.23%. For medium-size green spaces, for every 1/4 increase in the distance ratio, the humidifying intensity decreased by 3.02%. For large-size green spaces, for every 1/4 increase in the distance ratio, the humidifying intensity de-creased by 6.14%. For small, medium, and large green spaces, there was a positive correlation between canopy density and humidifying intensity. The positive correlation between canopy density and humidifying intensity in medium-size green spaces was extremely significant (R2 = 0.925). The greater the canopy density, the greater the humidifying intensity. For medium-size green spaces, for every 0.5 increase in canopy density, the humidifying intensity increased by 3.29%. For small-size green spaces, for every 0.5 increase in canopy density, the humidifying intensity increased by 3.17%. For large-size green spaces, for every 0.5 increase in canopy density, the humidifying intensity increased by 4.06% (Fig. 15).
    Figure 14Linear regressions between the distance from different types of green space to water area, canopy density and humidifying intensity.Full size imageFigure 15Correlation of different green space types with water distance, canopy density and cooling and humidifying intensity.Full size imageEffect of shape and area of water bodies on microclimatic effects based on numerical simulationBanded waterWe constructed a numerical simulation model to explore the effects of a simulated increase in water body area on cooling and humidification. Figure 16 shows the simulated distribution characteristics of temperature and relative humidity after a 5% and 10% increase in water area at 14:00 when temperatures were high. The results suggest that between 7:00 and 10:00, with a 5% and 10% increase in water area, the air temperature was basically the same and the cooling effect was insignificant. However, between 12:00 and 19:00 and particularly in the hours between 13:00 and 16:00 when temperatures were highest, a 5% increase in water area produced a significant cooling effect, with a daily average value of 0.05 °C and a maximum value of 0.09 °C. A 10% increase in water area produced an extremely significant cooling effect, with a daily average value of 0.07 °C and a maximum value of 0.14 °C. From 11:00 to 19:00, a 5% increase in water area produced a significant humidifying effect, with a daily average value of 0.08% and a maximum value of 0.17%. A 10% increase produced an extremely significant humidifying effect, with a daily average value of 0.13% and a maximum value of 0.26% (See supplementary file).Figure 16Distribution characteristics of cooling and humidifying effects of simulated increase of banded water area at 14:00. (a) original cooling effect of banded water in the sample area; (b) cooling effect of 5% increase in water area; (c) cooling effect of 10% increase in water area; (d) original humidifying effect of banded water in the sample area; (e) humidifying effect of 5% increase in water area; (f) humidifying effect of 10% increase of water area.Full size imageMassive waterFigure 17 shows the simulated distribution characteristics of the cooling and humidifying effects after a 5% and 10% increase in the water area at 14:00 when temperatures were high. Between 8:00 and 19:00, a 5% and 10% increase in water area produced a significant cooling effect. At 19:00, the numerical simulation result was abnormal when the water area increased by 5% and 10%; at 13:00, the numerical simulation result was also ab-normal when the water area increased by 10%. After excluding the abnormal simulated data, a 5% increase in water area produced a cooling effect, with a daily average value of 0.06 °C and a maximum value of 0.10 °C. A 10% increase in water area produced an extremely significant cooling effect, with a daily average value of 0.10 °C and a maximum value of 0.18 °C. Between 11:00 and 19:00, a 5% increase in water area produced a significant humidifying effect, with a daily average value of 0.05% and a maximum value of 0.13%. A 10% increase in water area produced an extremely significant humidifying effect, with a daily average value of 0.13% and a maximum value of 0.27% (See supplementary file).Figure 17Distribution characteristics of cooling and humidifying effects of simulated increase of massive water area at 14:00. (a) original cooling effect of massive water in the sample area; (b) cooling effect of 5% increase in water area; (c) cooling effect of 10% increase in water area; (d) original humidifying effect of massive water in the sample area; (e) humidifying effect of 5% increase in water area; (f) humidifying effect of 10% increase of water area.Full size imageAnnular waterFigure 18 shows the simulated distribution characteristics of the cooling and humidifying effects after a 5% and 10% increase in the area of the annular water body at 14:00 when temperatures were high. Between 7:00 and 19:00, a 5% and 10% increase in water area produced a significant cooling effect. Between 11:00 and 16:00 when temperatures were high, a 5% increase in water area produced a cooling effect, with a daily average value of 0.06 °C and a maximum value of 0.14 °C°C and a 10% increase in water area produced an extremely significant cooling effect, with a daily average value of 0.13 °C and a maximum value of 0.28 °C. Between 7:00 and 19:00, a 5% and 10% increase in water area produced significant humidifying effects. Between 11:00 and 16:00 when temperatures were high, a 5% increase in water area produced an extremely significant humidifying effect, with a daily average value of 0.17% and a maximum value of 0.39% and a 10% increase in water area produced an extremely significant humidifying effect with a daily average value of 0.38% and a maximum value of 0.81% (See supplementary file).Figure 18Distribution characteristics of cooling and humidifying effects of simulated increase of annular water area at 14:00. (a) original cooling effect of annular water in the sample area; (b) cooling effect of 5% increase in water area; (c) cooling effect of 10% increase in water area; (d) original humidifying effect of annular water in the sample area; (e) humidifying effect of 5% increase in water area; (f) humidifying effect of 10% increase of water area.Full size image More