{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T08:18:54Z","timestamp":1779178734550,"version":"3.51.4"},"reference-count":61,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2022,3,3]],"date-time":"2022-03-03T00:00:00Z","timestamp":1646265600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["U19AI062629"],"award-info":[{"award-number":["U19AI062629"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Peptidoglycan (PGN), a polymeric glycan macromolecule, is a major constituent of the bacterial cell wall and a conserved pathogen-associated molecular pattern (PAMP) that triggers immune responses through cytosolic sensors. Immune cells encounter both PGN polymers and hydrolyzed muropeptides during infections, and primary human innate immune cells respond better to polymeric PGN than the minimal bioactive subunit muramyl dipeptide (MDP). While MDP is internalized through macropinocytosis and\/or clathrin-mediated endocytosis, the internalization of particulate polymeric PGN is unresolved. We show here that PGN macromolecules isolated from Bacillus anthracis display a broad range of sizes, making them amenable for multiple internalization pathways. Pharmacologic inhibition indicates that PGN primarily, but not exclusively, is internalized by actin-dependent endocytosis. An alternate clathrin-independent but dynamin dependent pathway supports 20\u201330% of PGN uptake. In primary monocytes, this alternate pathway does not require activities of RhoA, Cdc42 or Arf6 small GTPases. Selective inhibition of PGN uptake shows that phagolysosomal trafficking, processing and downstream immune responses are drastically affected by actin depolymerization, while dynamin inhibition has a smaller effect. Overall, we show that polymeric PGN internalization occurs through two endocytic pathways with distinct potentials to trigger immune responses.<\/jats:p>","DOI":"10.3390\/microorganisms10030552","type":"journal-article","created":{"date-parts":[[2022,3,3]],"date-time":"2022-03-03T09:24:53Z","timestamp":1646299493000},"page":"552","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Internalization of Polymeric Bacterial Peptidoglycan Occurs through Either Actin or Dynamin Dependent Pathways"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4809-9885","authenticated-orcid":false,"given":"Narcis I.","family":"Popescu","sequence":"first","affiliation":[{"name":"Department of Arthritis and Clinical Immunology, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jackie","family":"Cochran","sequence":"additional","affiliation":[{"name":"Department of Arthritis and Clinical Immunology, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elizabeth","family":"Duggan","sequence":"additional","affiliation":[{"name":"Department of Arthritis and Clinical Immunology, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J\u0119drzej","family":"Kluza","sequence":"additional","affiliation":[{"name":"Department of Arthritis and Clinical Immunology, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert","family":"Silasi","sequence":"additional","affiliation":[{"name":"Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kenneth Mark","family":"Coggeshall","sequence":"additional","affiliation":[{"name":"Department of Arthritis and Clinical Immunology, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1007\/s00709-010-0164-2","article-title":"Hijacking the endocytic machinery by microbial pathogens","volume":"244","author":"Lin","year":"2010","journal-title":"Protoplasma"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1146\/annurev-pathol-011811-132445","article-title":"The cell biology of phagocytosis","volume":"7","author":"Flannagan","year":"2012","journal-title":"Annu. Rev. Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Lin, X.P., Mintern, J.D., and Gleeson, P.A. (2020). Macropinocytosis in Different Cell Types: Similarities and Differences. Membranes Basel, 10.","DOI":"10.3390\/membranes10080177"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1038\/nrm2216","article-title":"Pathways of clathrin-independent endocytosis","volume":"8","author":"Mayor","year":"2007","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.tcb.2012.10.007","article-title":"Targeting membrane trafficking in infection prophylaxis: Dynamin inhibitors","volume":"23","author":"Harper","year":"2013","journal-title":"Trends Cell Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1111\/j.1574-6976.2007.00094.x","article-title":"Peptidoglycan structure and architecture","volume":"32","author":"Vollmer","year":"2008","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"624","DOI":"10.1016\/j.cell.2016.05.076","article-title":"Hexokinase Is an Innate Immune Receptor for the Detection of Bacterial Peptidoglycan","volume":"166","author":"Wolf","year":"2016","journal-title":"Cell"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Langer, M., Girton, A.W., Popescu, N.I., Burgett, T., Metcalf, J.P., and Coggeshall, K.M. (2018). Neither Lys- and DAP-type peptidoglycans stimulate mouse or human innate immune cells via Toll-like receptor 2. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0193207"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"41702","DOI":"10.1074\/jbc.M307198200","article-title":"Peptidoglycan molecular requirements allowing detection by Nod1 and Nod2","volume":"278","author":"Girardin","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2423","DOI":"10.4049\/jimmunol.1201302","article-title":"Anti-peptidoglycan antibodies and Fcgamma receptors are the key mediators of inflammation in Gram-positive sepsis","volume":"189","author":"Sun","year":"2012","journal-title":"J. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e00076-18","DOI":"10.1128\/IAI.00076-18","article-title":"Serum Amyloid P and IgG Exhibit Differential Capabilities in the Activation of the Innate Immune System in Response to Bacillus anthracis Peptidoglycan","volume":"86","author":"Girton","year":"2018","journal-title":"Infect. Immun."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e2104347118","DOI":"10.1073\/pnas.2104347118","article-title":"Complement C5 inhibition protects against hemolytic anemia and acute kidney injury in anthrax peptidoglycan-induced sepsis in baboons","volume":"118","author":"Keshari","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1038\/nri3089","article-title":"Peptidoglycan recognition proteins: Modulators of the microbiome and inflammation","volume":"11","author":"Royet","year":"2011","journal-title":"Nat. Rev. Immunol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"702","DOI":"10.1038\/ni945","article-title":"An essential role for NOD1 in host recognition of bacterial peptidoglycan containing diaminopimelic acid","volume":"4","author":"Chamaillard","year":"2003","journal-title":"Nat. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1016\/j.molcel.2007.01.032","article-title":"Reconstituted NALP1 inflammasome reveals two-step mechanism of caspase-1 activation","volume":"25","author":"Faustin","year":"2007","journal-title":"Mol. Cell"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"849","DOI":"10.1182\/blood-2017-10-813618","article-title":"Peptidoglycan induces disseminated intravascular coagulation in baboons through activation of both coagulation pathways","volume":"132","author":"Popescu","year":"2018","journal-title":"Blood"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"13498","DOI":"10.1073\/pnas.1904170116","article-title":"Borrelia burgdorferi peptidoglycan is a persistent antigen in patients with Lyme arthritis","volume":"116","author":"Jutras","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1794","DOI":"10.1016\/S0140-6736(05)66582-8","article-title":"Muramyl dipeptide and toll-like receptor sensitivity in NOD2-associated Crohn\u2019s disease","volume":"365","author":"Ghosh","year":"2005","journal-title":"Lancet"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"766","DOI":"10.1038\/s41564-019-0381-1","article-title":"Antibody neutralization of microbiota-derived circulating peptidoglycan dampens inflammation and ameliorates autoimmunity","volume":"4","author":"Huang","year":"2019","journal-title":"Nat. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2418","DOI":"10.1128\/IAI.00170-10","article-title":"Inflammatory cytokine response to Bacillus anthracis peptidoglycan requires phagocytosis and lysosomal trafficking","volume":"78","author":"Iyer","year":"2010","journal-title":"Infect. Immun."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1110","DOI":"10.1016\/j.cell.2010.11.037","article-title":"Lipoprotein cofactors located in the outer membrane activate bacterial cell wall polymerases","volume":"143","author":"Markovski","year":"2010","journal-title":"Cell"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1097","DOI":"10.1016\/j.cell.2010.11.038","article-title":"Regulation of peptidoglycan synthesis by outer-membrane proteins","volume":"143","author":"Typas","year":"2010","journal-title":"Cell"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"500","DOI":"10.3389\/fmicb.2019.00500","article-title":"Peptidoglycan Muropeptides: Release, Perception, and Functions as Signaling Molecules","volume":"10","author":"Irazoki","year":"2019","journal-title":"Front. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4321","DOI":"10.4049\/jimmunol.0802197","article-title":"Clathrin- and dynamin-dependent endocytic pathway regulates muramyl dipeptide internalization and NOD2 activation","volume":"182","author":"Franchi","year":"2009","journal-title":"J. Immunol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1038\/nature13133","article-title":"Endosomes are specialized platforms for bacterial sensing and NOD2 signalling","volume":"509","author":"Nakamura","year":"2014","journal-title":"Nature"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1518","DOI":"10.1111\/cmi.12658","article-title":"Bacterial membrane vesicles: Biogenesis, immune regulation and pathogenesis","volume":"18","author":"Pathirana","year":"2016","journal-title":"Cell Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1111\/j.1462-5822.2009.01404.x","article-title":"Bacterial membrane vesicles deliver peptidoglycan to NOD1 in epithelial cells","volume":"12","author":"Kaparakis","year":"2010","journal-title":"Cell Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4034","DOI":"10.1128\/IAI.01980-14","article-title":"Aggregatibacter actinomycetemcomitans outer membrane vesicles are internalized in human host cells and trigger NOD1- and NOD2-dependent NF-kappaB activation","volume":"82","author":"Thay","year":"2014","journal-title":"Infect. Immun."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Mayer-Scholl, A., Hurwitz, R., Brinkmann, V., Schmid, M., Jungblut, P., Weinrauch, Y., and Zychlinsky, A. (2005). Human neutrophils kill Bacillus anthracis. PLoS Pathog., 1.","DOI":"10.1371\/journal.ppat.0010023"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"E6390","DOI":"10.1073\/pnas.1706818114","article-title":"Inhibition of complement C5 protects against organ failure and reduces mortality in a baboon model of Escherichia coli sepsis","volume":"114","author":"Keshari","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3841","DOI":"10.4049\/jimmunol.1004058","article-title":"Cutting edge: Primary innate immune cells respond efficiently to polymeric peptidoglycan, but not to peptidoglycan monomers","volume":"186","author":"Iyer","year":"2011","journal-title":"J. Immunol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2436","DOI":"10.1182\/bloodadvances.2019000513","article-title":"Monocyte procoagulant responses to anthrax peptidoglycan are reinforced by proinflammatory cytokine signaling","volume":"3","author":"Popescu","year":"2019","journal-title":"Blood Adv."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Popescu, N.I., Keshari, R.S., Cochran, J., Coggeshall, K.M., and Lupu, F. (2020). C3 Opsonization of Anthrax Bacterium and Peptidoglycan Supports Recognition and Activation of Neutrophils. Microorganisms, 8.","DOI":"10.3390\/microorganisms8071039"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Langer, M., Malykhin, A., Maeda, K., Chakrabarty, K., Williamson, K.S., Feasley, C.L., West, C.M., Metcalf, J.P., and Coggeshall, K.M. (2008). Bacillus anthracis peptidoglycan stimulates an inflammatory response in monocytes through the p38 mitogen-activated protein kinase pathway. PLoS ONE, 3.","DOI":"10.1371\/journal.pone.0003706"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1038\/s42003-019-0513-4","article-title":"A nanoscale, multi-parametric flow cytometry-based platform to study mitochondrial heterogeneity and mitochondrial DNA dynamics","volume":"2","author":"MacDonald","year":"2019","journal-title":"Commun. Biol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3986","DOI":"10.1091\/mbc.e11-04-0379","article-title":"A pharmacological cocktail for arresting actin dynamics in living cells","volume":"22","author":"Peng","year":"2011","journal-title":"Mol. Biol. Cell"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"641","DOI":"10.1038\/s41589-019-0262-1","article-title":"Disruption of endocytosis through chemical inhibition of clathrin heavy chain function","volume":"15","author":"Dejonghe","year":"2019","journal-title":"Nat. Chem. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"35966","DOI":"10.1074\/jbc.M111.283879","article-title":"Dynamin inhibition blocks botulinum neurotoxin type A endocytosis in neurons and delays botulism","volume":"286","author":"Harper","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3155","DOI":"10.1073\/pnas.1212324110","article-title":"Small-molecule inhibitors targeting G-protein-coupled Rho guanine nucleotide exchange factors","volume":"110","author":"Shang","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"4217","DOI":"10.1038\/s41467-018-06738-5","article-title":"Mechanochemical feedback control of dynamin independent endocytosis modulates membrane tension in adherent cells","volume":"9","author":"Thottacherry","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Karnik, R., Ludlow, M.J., Abuarab, N., Smith, A.J., Hardy, M.E., Elliott, D.J., and Sivaprasadarao, A. (2013). Endocytosis of HERG is clathrin-independent and involves arf6. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0085630"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"5125","DOI":"10.1021\/acs.biochem.7b00706","article-title":"Family-wide Analysis of the Inhibition of Arf Guanine Nucleotide Exchange Factors with Small Molecules: Evidence of Unique Inhibitory Profiles","volume":"56","author":"Benabdi","year":"2017","journal-title":"Biochemistry"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"8680","DOI":"10.1074\/jbc.273.15.8680","article-title":"Binding of bacterial peptidoglycan to CD14","volume":"273","author":"Dziarski","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1056\/NEJM199909093411107","article-title":"Anthrax","volume":"341","author":"Dixon","year":"1999","journal-title":"N. Engl. J. Med."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Popov, S.G. (2008). New Candidate Anthrax Pathogenic Factors, Springer.","DOI":"10.1007\/978-1-59745-569-5_4"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"914","DOI":"10.1111\/jcmm.12075","article-title":"The sepsis model: An emerging hypothesis for the lethality of inhalation anthrax","volume":"17","author":"Coggeshall","year":"2013","journal-title":"J. Cell Mol. Med."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1333","DOI":"10.1164\/rccm.201102-0209CI","article-title":"Anthrax infection","volume":"184","author":"Sweeney","year":"2011","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"978","DOI":"10.1093\/infdis\/jit247","article-title":"Bacillus anthracis cell wall peptidoglycan but not lethal or edema toxins produces changes consistent with disseminated intravascular coagulation in a rat model","volume":"208","author":"Qiu","year":"2013","journal-title":"J. Infect. Dis."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"571","DOI":"10.1182\/blood-2013-02-486613","article-title":"Bacillus anthracis peptidoglycan activates human platelets through FcgammaRII and complement","volume":"122","author":"Sun","year":"2013","journal-title":"Blood"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"11284","DOI":"10.1038\/ncomms11284","article-title":"Calcium-sensing receptors signal constitutive macropinocytosis and facilitate the uptake of NOD2 ligands in macrophages","volume":"7","author":"Canton","year":"2016","journal-title":"Nat. Commun."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"e02030-16","DOI":"10.1128\/mBio.02030-16","article-title":"Dissecting Bacterial Cell Wall Entry and Signaling in Eukaryotic Cells: An Actin-Dependent Pathway Parallels Platelet-Activating Factor Receptor-Mediated Endocytosis","volume":"8","author":"Loh","year":"2017","journal-title":"mBio"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1038\/nature14067","article-title":"Endophilin marks and controls a clathrin-independent endocytic pathway","volume":"517","author":"Boucrot","year":"2015","journal-title":"Nature"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1027","DOI":"10.1084\/jem.186.7.1027","article-title":"A critical role for Syk in signal transduction and phagocytosis mediated by Fcgamma receptors on macrophages","volume":"186","author":"Crowley","year":"1997","journal-title":"J. Exp. Med."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"4554","DOI":"10.1182\/blood-2005-09-3616","article-title":"Protein-tyrosine kinase Syk is required for pathogen engulfment in complement-mediated phagocytosis","volume":"107","author":"Shi","year":"2006","journal-title":"Blood"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1717","DOI":"10.1126\/science.282.5394.1717","article-title":"Identification of two distinct mechanisms of phagocytosis controlled by different Rho GTPases","volume":"282","author":"Caron","year":"1998","journal-title":"Science"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"473","DOI":"10.4049\/jimmunol.165.1.473","article-title":"Differential involvement of Src family kinases in Fc gamma receptor-mediated phagocytosis","volume":"165","author":"Suzuki","year":"2000","journal-title":"J. Immunol."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Abrami, L., Bischofberger, M., Kunz, B., Groux, R., and Van der Goot, F.G. (2010). Endocytosis of the anthrax toxin is mediated by clathrin, actin and unconventional adaptors. PLoS Pathog., 6.","DOI":"10.1371\/journal.ppat.1000792"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/S1471-4906(01)02161-5","article-title":"Caveolae and caveolin in immune cells: Distribution and functions","volume":"23","author":"Harris","year":"2002","journal-title":"Trends Immunol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"636","DOI":"10.1038\/39376","article-title":"Survival of FimH-expressing enterobacteria in macrophages relies on glycolipid traffic","volume":"389","author":"Baorto","year":"1997","journal-title":"Nature"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"5186","DOI":"10.4049\/jimmunol.165.9.5186","article-title":"Nonopsonic phagocytosis of Mycobacterium kansasii by human neutrophils depends on cholesterol and is mediated by CR3 associated with glycosylphosphatidylinositol-anchored proteins","volume":"165","author":"Peyron","year":"2000","journal-title":"J. Immunol."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"2090","DOI":"10.1091\/mbc.E15-11-0756","article-title":"Src-dependent phosphorylation of caveolin-1 Tyr-14 promotes swelling and release of caveolae","volume":"27","author":"Zimnicka","year":"2016","journal-title":"Mol. Biol. Cell"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/10\/3\/552\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:31:24Z","timestamp":1760135484000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/10\/3\/552"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,3]]},"references-count":61,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["microorganisms10030552"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms10030552","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,3]]}}}