{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,7]],"date-time":"2026-02-07T19:54:10Z","timestamp":1770494050165,"version":"3.49.0"},"reference-count":58,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2022,1,18]],"date-time":"2022-01-18T00:00:00Z","timestamp":1642464000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000027","name":"National Institute on Alcohol Abuse and Alcoholism","doi-asserted-by":"publisher","award":["R00-AA026336"],"award-info":[{"award-number":["R00-AA026336"]}],"id":[{"id":"10.13039\/100000027","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000738","name":"Department of Veterans Affairs","doi-asserted-by":"publisher","award":["IK6 BX003781"],"award-info":[{"award-number":["IK6 BX003781"]}],"id":[{"id":"10.13039\/100000738","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Pneumococcal pneumonia is a leading cause of morbidity and mortality worldwide. An increased susceptibility is due, in part, to compromised immune function. Zinc is required for proper immune function, and an insufficient dietary intake increases the risk of pneumonia. Our group was the first to reveal that the Zn transporter, ZIP8, is required for host defense. Furthermore, the gut microbiota that is essential for lung immunity is adversely impacted by a commonly occurring defective ZIP8 allele in humans. Taken together, we hypothesized that loss of the ZIP8 function would lead to intestinal dysbiosis and impaired host defense against pneumonia. To test this, we utilized a novel myeloid-specific Zip8KO mouse model in our studies. The comparison of the cecal microbial composition of wild-type and Zip8KO mice revealed significant differences in microbial community structure. Most strikingly, upon a S. pneumoniae lung infection, mice recolonized with Zip8KO-derived microbiota exhibited an increase in weight loss, bacterial dissemination, and lung inflammation compared to mice recolonized with WT microbiota. For the first time, we reveal the critical role of myeloid-specific ZIP8 on the maintenance of the gut microbiome structure, and that loss of ZIP8 leads to intestinal dysbiosis and impaired host defense in the lung. Given the high incidence of dietary Zn deficiency and the ZIP8 variant allele in the human population, additional investigation is warranted to improve surveillance and treatment strategies.<\/jats:p>","DOI":"10.3390\/ijms23031022","type":"journal-article","created":{"date-parts":[[2022,1,18]],"date-time":"2022-01-18T08:09:20Z","timestamp":1642493360000},"page":"1022","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["ZIP8-Mediated Intestinal Dysbiosis Impairs Pulmonary Host Defense against Bacterial Pneumonia"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5356-1413","authenticated-orcid":false,"given":"Derrick R.","family":"Samuelson","sequence":"first","affiliation":[{"name":"Department of Internal Medicine-Pulmonary Division, College of Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Deandra R.","family":"Smith","sequence":"additional","affiliation":[{"name":"Department of Pharmacy Practice and Science, College of Pharmacy, University of Nebraska Medical Center, Omaha, NE 68198, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kelly C.","family":"Cunningham","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine-Pulmonary Division, College of Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Todd A.","family":"Wyatt","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine-Pulmonary Division, College of Medicine, University of Nebraska Medical Center, Omaha, NE 68198, USA"},{"name":"Department of Environmental, Agricultural and Occupational Health, College of Public Health, University of Nebraska Medical Center, Omaha, NE 68198, USA"},{"name":"Veterans Affairs Nebraska-Western Iowa Health Care System, Omaha, NE 68105, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sannette C.","family":"Hall","sequence":"additional","affiliation":[{"name":"Department of Pharmacy Practice and Science, College of Pharmacy, University of Nebraska Medical Center, Omaha, NE 68198, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4169-5027","authenticated-orcid":false,"given":"Daryl J.","family":"Murry","sequence":"additional","affiliation":[{"name":"Department of Pharmacy Practice and Science, College of Pharmacy, University of Nebraska Medical Center, Omaha, NE 68198, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6455-5388","authenticated-orcid":false,"given":"Yashpal S.","family":"Chhonker","sequence":"additional","affiliation":[{"name":"Department of Pharmacy Practice and Science, College of Pharmacy, University of Nebraska Medical Center, Omaha, NE 68198, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9977-2857","authenticated-orcid":false,"given":"Daren L.","family":"Knoell","sequence":"additional","affiliation":[{"name":"Department of Pharmacy Practice and Science, College of Pharmacy, University of Nebraska Medical Center, Omaha, NE 68198, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1395","DOI":"10.1086\/657313","article-title":"Integrated analysis of FOCUS 1 and FOCUS 2: Randomized, doubled-blinded, multicenter phase 3 trials of the efficacy and safety of ceftaroline fosamil versus ceftriaxone in patients with community-acquired pneumonia","volume":"51","author":"File","year":"2010","journal-title":"Clin. Infect. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.jinf.2013.03.003","article-title":"Can an etiologic agent be identified in adults who are hospitalized for community-acquired pneumonia: Results of a one-year study","volume":"67","author":"Musher","year":"2013","journal-title":"J. Infect."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"610","DOI":"10.1378\/chest.07-1456","article-title":"A comparative study of community-acquired pneumonia patients admitted to the ward and the ICU","volume":"133","author":"Restrepo","year":"2008","journal-title":"Chest"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1813","DOI":"10.1093\/infdis\/jit506","article-title":"Distribution of 13-valent pneumococcal conjugate vaccine Streptococcus pneumoniae serotypes in US adults aged \u226550 years with community-acquired pneumonia","volume":"208","author":"Sherwin","year":"2013","journal-title":"J. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"766","DOI":"10.1164\/ajrccm.165.6.2103038","article-title":"Hospitalized community-acquired pneumonia in the elderly: Age- and sex-related patterns of care and outcome in the United States","volume":"165","author":"Kaplan","year":"2002","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_6","first-page":"350","article-title":"Prospective study of pneumonia hospitalizations and mortality of U.S. older people: The role of chronic conditions, health behaviors, and nutritional status","volume":"104","author":"LaCroix","year":"1989","journal-title":"Public Health Rep."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1007\/s10522-015-9614-8","article-title":"Immune ageing and susceptibility to Streptococcus pneumoniae","volume":"17","author":"Goncalves","year":"2016","journal-title":"Biogerontology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1407S","DOI":"10.1093\/jn\/130.5.1407S","article-title":"Zinc-altered immune function and cytokine production","volume":"130","author":"Rink","year":"2000","journal-title":"J. Nutr."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1399S","DOI":"10.1093\/jn\/130.5.1399S","article-title":"The dynamic link between the integrity of the immune system and zinc status","volume":"130","author":"Fraker","year":"2000","journal-title":"J. Nutr."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1452S","DOI":"10.1093\/jn\/133.5.1452S","article-title":"Zinc-altered immune function","volume":"133","author":"Ibs","year":"2003","journal-title":"J. Nutr."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1146\/annurev.nutr.24.012003.132454","article-title":"Reprogramming of the immune system during zinc deficiency","volume":"24","author":"Fraker","year":"2004","journal-title":"Annu. Rev. Nutr."},{"key":"ref_12","first-page":"E1002","article-title":"Changes in cytokine production and T cell subpopulations in experimentally induced zinc-deficient humans","volume":"272","author":"Beck","year":"1997","journal-title":"Am. J. Physiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1146\/annurev.nutr.23.011702.073054","article-title":"Zinc and the risk for infectious disease","volume":"24","author":"Black","year":"2004","journal-title":"Annu. Rev. Nutr."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/S0047-6374(00)00194-9","article-title":"Zinc, infections and immunosenescence","volume":"121","author":"Mocchegiani","year":"2000","journal-title":"Mech. Ageing Dev."},{"key":"ref_15","first-page":"94","article-title":"Zinc nutriture and cell-mediated immunity in the aged","volume":"53","author":"Wagner","year":"1983","journal-title":"Int. J. Vitam. Nutr. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1111\/j.1753-4887.2009.00253.x","article-title":"Low zinc status: A new risk factor for pneumonia in the elderly?","volume":"68","author":"Barnett","year":"2010","journal-title":"Nutr. Rev."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1093\/ajcn\/85.3.837","article-title":"Zinc supplementation decreases incidence of infections in the elderly: Effect of zinc on generation of cytokines and oxidative stress","volume":"85","author":"Prasad","year":"2007","journal-title":"Am. J. Clin. Nutr."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1001\/archinte.159.7.748","article-title":"Impact of trace elements and vitamin supplementation on immunity and infections in institutionalized elderly patients: A randomized controlled trial. MIN. VIT. AOX. geriatric network","volume":"159","author":"Girodon","year":"1999","journal-title":"Arch. Intern. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1159\/000177984","article-title":"Effect of micronutrient supplementation on infection in institutionalized elderly subjects: A controlled trial","volume":"41","author":"Girodon","year":"1997","journal-title":"Ann. Nutr. Metab."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1016\/j.cmet.2017.05.008","article-title":"Understanding the Holobiont: How Microbial Metabolites Affect Human Health and Shape the Immune System","volume":"26","author":"Postler","year":"2017","journal-title":"Cell Metab."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1085","DOI":"10.3389\/fmicb.2015.01085","article-title":"Regulation of lung immunity and host defense by the intestinal microbiota","volume":"6","author":"Samuelson","year":"2015","journal-title":"Front. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1016\/j.celrep.2013.01.009","article-title":"ZIP8 regulates host defense through zinc-mediated inhibition of NF-kappaB","volume":"3","author":"Liu","year":"2013","journal-title":"Cell Rep."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Pyle, C.J., Akhter, S., Bao, S., Dodd, C.E., Schlesinger, L.S., and Knoell, D.L. (2017). Zinc Modulates Endotoxin-Induced Human Macrophage Inflammation through ZIP8 Induction and C\/EBPbeta Inhibition. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0169531"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Pyle, C.J., Azad, A.K., Papp, A.C., Sadee, W., Knoell, D.L., and Schlesinger, L.S. (2017). Elemental Ingredients in the Macrophage Cocktail: Role of ZIP8 in Host Response to Mycobacterium tuberculosis. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18112375"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1038\/ng.3570","article-title":"Detection and interpretation of shared genetic influences on 42 human traits","volume":"48","author":"Pickrell","year":"2016","journal-title":"Nat. Genet."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1002\/ajmg.b.32545","article-title":"The highly pleiotropic gene SLC39A8 as an opportunity to gain insight into the molecular pathogenesis of schizophrenia","volume":"177","author":"Costas","year":"2018","journal-title":"Am. J. Med. Genet. B Neuropsychiatr. Genet."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"125","DOI":"10.3389\/fgene.2014.00125","article-title":"Genome wide association study of SNP-, gene-, and pathway-based approaches to identify genes influencing susceptibility to Staphylococcus aureus infections","volume":"5","author":"Ye","year":"2014","journal-title":"Front. Genet."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1097\/MCO.0b013e32833eebe5","article-title":"Butyrate and other short-chain fatty acids as modulators of immunity: What relevance for health?","volume":"13","author":"Meijer","year":"2010","journal-title":"Curr. Opin. Clin. Nutr. Metab. Care"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Sin, D.D., Leung, R., Gan, W.Q., and Man, S.P. (2007). Circulating surfactant protein D as a potential lung-specific biomarker of health outcomes in COPD: A pilot study. BMC Pulm. Med., 7.","DOI":"10.1186\/1471-2466-7-13"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1177\/0267659114566063","article-title":"Prediction of reversibility of intestinal mucosal damage after ischemia-reperfusion injury by plasma intestinal fatty acid-binding protein levels in pigs","volume":"30","author":"Kano","year":"2015","journal-title":"Perfusion"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Schurink, M., Kooi, E.M., Hulzebos, C.V., Kox, R.G., Groen, H., Heineman, E., Bos, A.F., and Hulscher, J.B.F. (2015). Intestinal fatty acid-binding protein as a diagnostic marker for complicated and uncomplicated necrotizing enterocolitis: A prospective cohort study. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0121336"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e00049","DOI":"10.1042\/BSR20130014","article-title":"Metal ions in macrophage antimicrobial pathways: Emerging roles for zinc and copper","volume":"33","author":"Stafford","year":"2013","journal-title":"Biosci. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1016\/j.tube.2011.12.006","article-title":"Resistance mechanisms of Mycobacterium tuberculosis against phagosomal copper overload","volume":"92","author":"Rowland","year":"2012","journal-title":"Tuberculosis"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1016\/S0968-0004(99)01386-9","article-title":"Iron overload: Molecular clues to its cause","volume":"24","author":"Kuhn","year":"1999","journal-title":"Trends Biochem. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1691","DOI":"10.1111\/j.1462-5822.2010.01529.x","article-title":"The struggle for iron\u2014A metal at the host-pathogen interface","volume":"12","author":"Nairz","year":"2010","journal-title":"Cell. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1146\/annurev-nutr-033009-083312","article-title":"Mammalian zinc transporters: Nutritional and physiologic regulation","volume":"29","author":"Lichten","year":"2009","journal-title":"Annu. Rev. Nutr."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1357","DOI":"10.4049\/jimmunol.2001395","article-title":"Critical Role of Zinc Transporter (ZIP8) in Myeloid Innate Immune Cell Function and the Host Response against Bacterial Pneumonia","volume":"207","author":"Hall","year":"2021","journal-title":"J. Immunol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1007\/s10534-020-00279-5","article-title":"Effects of zinc status on age-related T cell dysfunction and chronic inflammation","volume":"34","author":"Wong","year":"2021","journal-title":"Biometals"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"416","DOI":"10.1007\/s10875-009-9298-4","article-title":"NK and NKT cells in aging and longevity: Role of zinc and metallothioneins","volume":"29","author":"Mocchegiani","year":"2009","journal-title":"J. Clin. Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1016\/j.exger.2007.11.006","article-title":"TH1 and TH2 cell polarization increases with aging and is modulated by zinc supplementation","volume":"43","author":"Uciechowski","year":"2008","journal-title":"Exp. Gerontol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2090","DOI":"10.1126\/science.1104742","article-title":"Hepcidin regulates cellular iron efflux by binding to ferroportin and inducing its internalization","volume":"306","author":"Nemeth","year":"2004","journal-title":"Science"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1411","DOI":"10.4049\/jimmunol.1101682","article-title":"Transient TLR activation restores inflammatory response and ability to control pulmonary bacterial infection in germfree mice","volume":"188","author":"Fagundes","year":"2012","journal-title":"J. Immunol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1512","DOI":"10.1038\/s41467-017-01803-x","article-title":"The microbiota protects against respiratory infection via GM-CSF signaling","volume":"8","author":"Brown","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"5354","DOI":"10.1073\/pnas.1019378108","article-title":"Microbiota regulates immune defense against respiratory tract influenza A virus infection","volume":"108","author":"Ichinohe","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"3273","DOI":"10.1038\/s41467-019-11152-6","article-title":"Microbiota-derived acetate protects against respiratory syncytial virus infection through a GPR43-type 1 interferon response","volume":"10","author":"Antunes","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Samuelson, D.R., Shellito, J.E., Maffei, V.J., Tague, E.D., Campagna, S.R., Blanchard, E.E., Luo, M., Taylor, C.M., Ronis, M.J.J., and Molina, P.E. (2017). Alcohol-associated intestinal dysbiosis impairs pulmonary host defense against Klebsiella pneumoniae. PLoS Pathog., 13.","DOI":"10.1371\/journal.ppat.1006426"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"3111","DOI":"10.1128\/IAI.70.6.3111-3121.2002","article-title":"Oral vaccination with subunit vaccines protects animals against aerosol infection with Mycobacterium tuberculosis","volume":"70","author":"Doherty","year":"2002","journal-title":"Infect. Immun."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"3781","DOI":"10.1016\/j.vaccine.2007.02.014","article-title":"Oral immunization of mice with the live vaccine strain (LVS) of Francisella tularensis protects mice against respiratory challenge with virulent type A F. tularensis","volume":"25","author":"KuoLee","year":"2007","journal-title":"Vaccine"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2655","DOI":"10.4049\/jimmunol.1502004","article-title":"Oral Immunization of Mice with Live Pneumocystis murina Protects against Pneumocystis Pneumonia","volume":"196","author":"Samuelson","year":"2016","journal-title":"J. Immunol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"114729","DOI":"10.1016\/j.jep.2021.114729","article-title":"Yanning Syrup ameliorates the lipopolysaccharide-induced inflammation: Adjusting the gut microbiota, short-chain fatty acids, and the CD4(+) T cell balance","volume":"283","author":"Wei","year":"2022","journal-title":"J. Ethnopharmacol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"e0018821","DOI":"10.1128\/IAI.00188-21","article-title":"Short-Chain Fatty Acids as a Potential Treatment for Infections: A Closer Look at the Lungs","volume":"89","author":"Machado","year":"2021","journal-title":"Infect. Immun."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"2934","DOI":"10.1016\/j.celrep.2020.02.013","article-title":"Gut Dysbiosis during Influenza Contributes to Pulmonary Pneumococcal Superinfection through Altered Short-Chain Fatty Acid Production","volume":"30","author":"Sencio","year":"2020","journal-title":"Cell Rep."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1038\/nmeth.3869","article-title":"DADA2: High-resolution sample inference from Illumina amplicon data","volume":"13","author":"Callahan","year":"2016","journal-title":"Nat. Methods"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"3123","DOI":"10.1093\/bioinformatics\/btu494","article-title":"STAMP: Statistical analysis of taxonomic and functional profiles","volume":"30","author":"Parks","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2","volume":"15","author":"Love","year":"2014","journal-title":"Genome Biol."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"McMurdie, P.J., and Holmes, S. (2013). phyloseq: An R package for reproducible interactive analysis and graphics of microbiome census data. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0061217"},{"key":"ref_57","unstructured":"Oksanen, J., Blanchet, F.G., Friendly, M., Kindt, R., Legendre, P., McGlinn, D., Minchin, P.R., O\u2019Hara, R.B., Simpson, G.L., and Solymos, P. (2021, December 27). vegan: Community Ecology Packag. R Package Version 2.5-7. Available online: https:\/\/cran.r-project.org\/package=vegan."},{"key":"ref_58","first-page":"672295","article-title":"PICRUSt2: An improved and extensible approach for metagenome inference","volume":"2019","author":"Douglas","year":"2019","journal-title":"BioRxiv"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/3\/1022\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:03:09Z","timestamp":1760133789000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/3\/1022"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,18]]},"references-count":58,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["ijms23031022"],"URL":"https:\/\/doi.org\/10.3390\/ijms23031022","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1,18]]}}}