{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T19:57:56Z","timestamp":1778702276497,"version":"3.51.4"},"reference-count":53,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2024,6,5]],"date-time":"2024-06-05T00:00:00Z","timestamp":1717545600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"South African Medical Research Council (self-initiated)","award":["9581"],"award-info":[{"award-number":["9581"]}]},{"name":"South African Medical Research Council (self-initiated)","award":["21\/76"],"award-info":[{"award-number":["21\/76"]}]},{"name":"Poliomyelitis Research Foundation","award":["9581"],"award-info":[{"award-number":["9581"]}]},{"name":"Poliomyelitis Research Foundation","award":["21\/76"],"award-info":[{"award-number":["21\/76"]}]},{"name":"The UKZN Research Office","award":["9581"],"award-info":[{"award-number":["9581"]}]},{"name":"The UKZN Research Office","award":["21\/76"],"award-info":[{"award-number":["21\/76"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Developing new anti-human immunodeficiency virus (HIV) drug candidates that target different sites in HIV-1 replication, with better resistance profiles and lower drug toxicity, is essential to eradicating HIV. This study investigated the potential of fractionated crude extracts of Alternaria alternata as immunomodulatory or anti-HIV drug candidates. Solid-phase extraction (SPE) was used to fractionate A. alternata PO4PR2 using three different columns: MAX (Mixed-mode, strong Anion-eXchange), MCX (Mixed-mode, strong Cation-eXchange), and HLB (Hydrophilic\u2013Lipophilic Balance) with methanol gradient methods (5%, 45%, and 95%). An MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay was used to assess the cell viability and cytotoxicity of the fractionated crude extract A. alternata PO4PR2 in the TZM-bl cell lines. This was followed by a luciferase-based antiviral assay to assess the antiviral activity of A. alternata PO4PR2. A time of addition (TOA) assay was performed to ascertain the mechanism of inhibition employed by the fractionated crude extract of A. alternata PO4PR2 in the HIV life cycle. The p24 titer was determined using an ELISA, while a luciferase-based antiviral assay was used to evaluate the HIV percentage inhibition for different HIV-1 replication cycles. The TOA assay was established using antiviral drugs that target different sites in the HIV replication cycle. These included maraviroc, azidothymidine, raltegravir, and amprenavir. The immunomodulatory effect of the fractionated crude extracts on CD4+ T cells was measured by a flow cytometric analysis, for which fluorochrome-labelled monoclonal antibodies were used as markers for activation (CD38 and HLA-DR) and exhaustion (PD-1). The MCX fraction demonstrated a more significant anti-HIV inhibition than that of the fractions generated in other columns, with an IC50 of 0.3619 \u00b5g\/mL, an HIV inhibition of 77%, 5% HLB (IC50: 0.7232 \u00b5g\/mL; HIV inhibition of 64%), and 5% MAX (IC50: 5.240 \u00b5g\/mL; HIV inhibition of 67%). It was evident from the time of addition data that the crude extract and the 5% MCX fraction inhibited viral binding (68%), reverse transcription (75%), integration (98%), and proteolysis (77%). It was shown that A. alternata (the MCX fraction) have a significant inhibitory effect on reverse transcription (75% HIV inhibition) and integration (100% HIV inhibition). The 5% MCX (p = 0.0062), 5% HLB (p = 0.0269), and 5% MAX (p = 0.0117) fractionated A. alternata crude extracts had low levels of CD4+ T cell (CD38 + HLA-DR+) activation compared to those of the AZT treatment, while CD4+ T cell activation was insignificant. The 5% MAX and HLB A. alternata fractions may possess immunomodulatory compounds with less anti-HIV-1 activity. A. alternata could be a key source of innovative anti-HIV drugs with immunomodulatory characteristics.<\/jats:p>","DOI":"10.3390\/microorganisms12061150","type":"journal-article","created":{"date-parts":[[2024,6,5]],"date-time":"2024-06-05T10:05:50Z","timestamp":1717581950000},"page":"1150","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Anti-HIV Activity and Immunomodulatory Properties of Fractionated Crude Extracts of Alternaria alternata"],"prefix":"10.3390","volume":"12","author":[{"given":"Mbali X.","family":"Kubheka","sequence":"first","affiliation":[{"name":"HIV Pathogenesis Programme, Doris Duke Medical Research Institute, School of Laboratory Medicine and Medical Sciences, University of KwaZulu-Natal, Durban 4041, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2323-282X","authenticated-orcid":false,"given":"Sizwe I.","family":"Ndlovu","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Food Technology, Faculty of Science, University of Johannesburg, Johannesburg 2092, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0808-9201","authenticated-orcid":false,"given":"Nompumelelo P.","family":"Mkhwanazi","sequence":"additional","affiliation":[{"name":"HIV Pathogenesis Programme, Doris Duke Medical Research Institute, School of Laboratory Medicine and Medical Sciences, University of KwaZulu-Natal, Durban 4041, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,6,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"624","DOI":"10.1038\/nrd1151","article-title":"Toxicity of antiretroviral therapy and implications for drug development","volume":"2","author":"Carr","year":"2003","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1007\/s40121-018-0201-6","article-title":"A Review of Long-Term Toxicity of Antiretroviral Treatment Regimens and Implications for an Aging Population","volume":"7","author":"Chawla","year":"2018","journal-title":"Infect. Dis. Ther."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"288","DOI":"10.4103\/0253-7613.96296","article-title":"Antiretroviral drugs: Critical issues and recent advances","volume":"44","author":"Desai","year":"2012","journal-title":"Indian J. Pharmacol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1016\/S1286-4579(03)00073-X","article-title":"Endophytes as sources of bioactive products","volume":"5","author":"Strobel","year":"2003","journal-title":"Microbes Infect."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Bacon, C.W., and White Jr, J.F. (2000). Physiological adaptations in the evolution of endophytism in the Clavicipitaceae. Microbial Endophytes, CRC Press.","DOI":"10.1201\/9781482277302-13"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1007\/s12013-013-9695-4","article-title":"Biological Evaluation of Endophytic Fungus, Chaetomium globosum JN711454, as Potential Candidate for Improving Drug Discovery","volume":"68","author":"Selim","year":"2014","journal-title":"Cell Biochem. Biophys."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1007\/s10295-003-0101-x","article-title":"Isolation, structure, and HIV-1-integrase inhibitory activity of structurally diverse fungal metabolites","volume":"30","author":"Singh","year":"2003","journal-title":"J. Ind. Microbiol. Biotechnol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1021\/np058128n","article-title":"Natural products from plant-associated microorganisms: Distribution, structural diversity, bioactivity, and implications of their occurrence","volume":"69","author":"Gunatilaka","year":"2006","journal-title":"J. Nat. Prod."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1515\/bot-2011-0095","article-title":"Antioxidants in mangrove plants and endophytic fungal associations","volume":"55","author":"Ravindran","year":"2012","journal-title":"Bot. Mar."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1016\/j.scitotenv.2018.06.213","article-title":"Developmental toxicity of synthetic phenolic antioxidants to the early life stage of zebrafish","volume":"643","author":"Yang","year":"2018","journal-title":"Sci. Total Environ."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Hoque, N., Khan, Z.R., Rashid, P.T., Begum, M.N., Sharmin, S., Hossain, M.J., Rana, M.S., and Sohrab, M.H. (2023). Antimicrobial, antioxidant, and cytotoxic properties of endophytic fungi isolated from Thysanolaena maxima Roxb., Dracaena spicata Roxb. and Aglaonema hookerianum Schott. BMC Complement. Med. Ther., 23.","DOI":"10.1186\/s12906-023-04185-4"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"6685","DOI":"10.1016\/j.bmcl.2010.09.010","article-title":"Xiamycin, a pentacyclic indolosesquiterpene with selective anti-HIV activity from a bacterial mangrove endophyte","volume":"20","author":"Ding","year":"2010","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Selim, K.A., Elkhateeb, W.A., Tawila, A.M., El-Beih, A.A., Abdel-Rahman, T.M., El-Diwany, A.I., and Ahmed, E.F. (2018). Antiviral and Antioxidant Potential of Fungal Endophytes of Egyptian Medicinal Plants. Fermentation, 4.","DOI":"10.3390\/fermentation4030049"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1111\/hiv.13116","article-title":"Bioactive phytocompound mulberroside C and endophytes of Morus alba as potential inhibitors of HIV-1 replication: A mechanistic evaluation","volume":"22","author":"Vora","year":"2021","journal-title":"HIV Med."},{"key":"ref_15","first-page":"5051","article-title":"Inhibition of HIV-1 by curcumin A, a novel curcumin analog","volume":"9","author":"Kumari","year":"2015","journal-title":"Drug Des. Dev. Ther."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1165","DOI":"10.1016\/0006-2952(95)98514-A","article-title":"Inhibition of human immunodeficiency virus type-1 integrase by curcumin","volume":"49","author":"Mazumder","year":"1995","journal-title":"Biochem. Pharmacol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3364","DOI":"10.1016\/j.bmcl.2005.05.032","article-title":"Active site binding modes of curcumin in HIV-1 protease and integrase","volume":"15","author":"Vajragupta","year":"2005","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"27539","DOI":"10.1038\/srep27539","article-title":"Curcumin inhibits HIV-1 by promoting Tat protein degradation","volume":"6","author":"Ali","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"6112","DOI":"10.1016\/j.bmc.2014.08.039","article-title":"Altertoxins with potent anti-HIV activity from Alternaria tenuissima QUE1Se, a fungal endophyte of Quercus emoryi","volume":"22","author":"Bashyal","year":"2014","journal-title":"Bioorg. Med. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"321","DOI":"10.4236\/pp.2015.67034","article-title":"In vitro anti-HIV activity of partially purified coumarin (s) isolated from fungal endophyte, Alternaria species of Calophyllum inophyllum","volume":"6","author":"Govindappa","year":"2015","journal-title":"Pharmacol. Pharm."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"104863","DOI":"10.1016\/j.fitote.2021.104863","article-title":"Coumarin-based derivatives with potential anti-HIV activity","volume":"150","author":"Xu","year":"2021","journal-title":"Fitoterapia"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Sharapov, A.D., Fatykhov, R.F., Khalymbadzha, I.A., Zyryanov, G.V., Chupakhin, O.N., and Tsurkan, M.V. (2023). Plant Coumarins with Anti-HIV Activity: Isolation and Mechanisms of Action. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24032839"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Nahar, L., Talukdar, A.D., Nath, D., Nath, S., Mehan, A., Ismail, F.M.D., and Sarker, S.D. (2020). Naturally Occurring Calanolides: Occurrence, Biosynthesis, and Pharmacological Properties Including Therapeutic Potential. Molecules, 25.","DOI":"10.3390\/molecules25214983"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Khalymbadzha, I.A., Fatykhov, R.F., Butorin, I.I., Sharapov, A.D., Potapova, A.P., Muthipeedika, N.J., Zyryanov, G.V., Melekhin, V.V., Tokhtueva, M.D., and Deev, S.L. (2024). Bioinspired Pyrano[2,3-f]chromen-8-ones: Ring C-Opened Analogues of Calanolide A: Synthesis and Anti-HIV-1 Evaluation. Biomimetics, 9.","DOI":"10.3390\/biomimetics9010044"},{"key":"ref_25","first-page":"5546","article-title":"Mechanistic insight into immunomodulatory effects of food-functioned plant secondary metabolites","volume":"63","author":"Nasrin","year":"2021","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.aspen.2014.11.004","article-title":"Studies on immunomodulatory effect of endophytic fungus Alternaria alternata on Spodoptera litura","volume":"18","author":"Kaur","year":"2015","journal-title":"J. Asia-Pac. Entomol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Abood, W.N., Fahmi, I., Abdulla, M.A., and Ismail, S. (2014). Immunomodulatory effect of an isolated fraction from Tinospora crispa on intracellular expression of INF-\u03b3, IL-6 and IL-8. BMC Complement. Altern. Med., 14.","DOI":"10.1186\/1472-6882-14-205"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Suwannarach, N., Kumla, J., Sujarit, K., Pattananandecha, T., Saenjum, C., and Lumyong, S. (2020). Natural Bioactive Compounds from Fungi as Potential Candidates for Protease Inhibitors and Immunomodulators to Apply for Coronaviruses. Molecules, 25.","DOI":"10.3390\/molecules25081800"},{"key":"ref_29","first-page":"19","article-title":"Antimicrobial and immunomodulatory activities of secondary metabolites of an endophytic fungus isolated from Ageratum conyzoides","volume":"5","author":"Ujam","year":"2019","journal-title":"Life Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1077159","DOI":"10.3389\/fgene.2022.1077159","article-title":"Secondary metabolites produced by endophytic fungi, Alternaria alternata, as potential inhibitors of the human immunodeficiency virus","volume":"13","author":"Nzimande","year":"2022","journal-title":"Front. Genet."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"6166","DOI":"10.1016\/j.bmcl.2011.07.123","article-title":"Microwave assisted synthesis and antimicrobial evaluation of new fused pyran derivatives bearing 2-morpholinoquinoline nucleus","volume":"21","author":"Makawana","year":"2011","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e202200020","DOI":"10.1002\/cmdc.202200020","article-title":"Cyclobutanes in small-molecule drug candidates","volume":"17","author":"Janssen","year":"2022","journal-title":"ChemMedChem"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1590\/fst.15619","article-title":"In vitro potential activity of some seaweeds as antioxidants and inhibitors of diabetic enzymes","volume":"40","author":"Ismail","year":"2019","journal-title":"Food Sci. Technol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"eaba6617","DOI":"10.1126\/sciadv.aba6617","article-title":"Gliotoxin, identified from a screen of fungal metabolites, disrupts 7SK snRNP, releases P-TEFb, and reverses HIV-1 latency","volume":"6","author":"Stoszko","year":"2020","journal-title":"Sci. Adv."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Makhwitine, J.P., Kumalo, H.M., Ndlovu, S.I., and Mkhwanazi, N.P. (2023). Epigenetic Induction of Secondary Metabolites Production in Endophytic Fungi Penicillium chrysogenum and GC-MS Analysis of Crude Metabolites with Anti-HIV-1 Activity. Microorganisms, 11.","DOI":"10.3390\/microorganisms11061404"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Fanele, A., and Ndlovu, S.I. (2023). Endophytic fungal species Nigrospora oryzae and Alternaria alternata exhibit antimicrobial activity against gram-positive and gram-negative multi-drug resistant clinical bacterial isolates. BMC Complement. Med. Ther., 23.","DOI":"10.1186\/s12906-023-04157-8"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1016\/j.virol.2005.06.003","article-title":"HIV-1 envelope pseudotyped viral vectors and infectious molecular clones expressing the same envelope glycoprotein have a similar neutralization phenotype, but culture in peripheral blood mononuclear cells is associated with decreased neutralization sensitivity","volume":"339","author":"Louder","year":"2005","journal-title":"Virology"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/j.jim.2013.11.022","article-title":"Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1","volume":"409","author":"Bailer","year":"2014","journal-title":"J. Immunol. Methods"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Cherne, M.D., Hall, J., Kellner, A., Chong, C.F., Cole, A.L., and Cole, A.M. (2019). Avirulins, a Novel Class of HIV-1 Reverse Transcriptase Inhibitors Effective in the Female Reproductive Tract Mucosa. Viruses, 11.","DOI":"10.3390\/v11050408"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"925","DOI":"10.1038\/nprot.2011.330","article-title":"A time-of\u2013drug addition approach to target identification of antiviral compounds","volume":"6","author":"Daelemans","year":"2011","journal-title":"Nat. Protoc."},{"key":"ref_41","first-page":"00017","article-title":"Luciferase Time-based, High-throughput Screening Assay for the Discovery of HIV-1 Inhibitors","volume":"1","author":"Lara","year":"2014","journal-title":"J. Hum. Virol. Retrovirol."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Chachage, M., Podola, L., Clowes, P., Nsojo, A., Bauer, A., Mgaya, O., Kowour, D., Froeschl, G., Maboko, L., and Hoelscher, M. (2014). Helminth-associated systemic immune activation and HIV co-receptor expression: Response to albendazole\/praziquantel treatment. PLoS Neglected Trop. Dis., 8.","DOI":"10.1371\/journal.pntd.0002755"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2865","DOI":"10.1128\/AAC.46.9.2865-2871.2002","article-title":"Immunomodulatory effect of zidovudine (ZDV) on cytotoxic T lymphocytes previously exposed to ZDV","volume":"46","author":"Francke","year":"2002","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1007\/s00228-015-1832-0","article-title":"Narrow therapeutic index drugs: A clinical pharmacological consideration to flecainide","volume":"71","author":"Tamargo","year":"2015","journal-title":"Eur. J. Clin. Pharmacol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"751","DOI":"10.1038\/nrd3801","article-title":"The determination and interpretation of the therapeutic index in drug development","volume":"11","author":"Muller","year":"2012","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1636","DOI":"10.1093\/cid\/ciz442","article-title":"Antiretroviral Therapy Reduces T-cell Activation and Immune Exhaustion Markers in Human Immunodeficiency Virus Controllers","volume":"70","author":"Li","year":"2019","journal-title":"Clin. Infect. Dis."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"S51","DOI":"10.1093\/infdis\/jiw287","article-title":"Cytokines and T-Cell Homeostasis in HIV Infection","volume":"214","author":"Freeman","year":"2016","journal-title":"J. Infect. Dis."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Schultz, H.S., Reedtz-Runge, S.L., B\u00e4ckstr\u00f6m, B.T., Lamberth, K., Pedersen, C.R., and Kvarnhammar, A.M. (2017). Quantitative analysis of the CD4+ T cell response to therapeutic antibodies in healthy donors using a novel T cell:PBMC assay. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0178544"},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Qiu, C., Wang, J., Zhu, L., Cheng, X., Xia, B., Jin, Y., Qin, R., Zhang, L., Hu, H., and Yan, J. (2022). Improving the ex vivo expansion of human tumor-reactive CD8 + T cells by targeting toll-like receptors. Front. Bioeng. Biotechnol., 10.","DOI":"10.3389\/fbioe.2022.1027619"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"2665","DOI":"10.4049\/jimmunol.1101165","article-title":"Functional exhaustion of CD4+ T lymphocytes during primary cytomegalovirus infection","volume":"189","author":"Antoine","year":"2012","journal-title":"J. Immunol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1111\/imr.12823","article-title":"T-cell exhaustion in HIV infection","volume":"292","author":"Fenwick","year":"2019","journal-title":"Immunol. Rev."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1038\/s41423-020-0402-2","article-title":"Functional exhaustion of antiviral lymphocytes in COVID-19 patients","volume":"17","author":"Zheng","year":"2020","journal-title":"Cell. Mol. Immunol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1016\/j.chom.2012.05.003","article-title":"Timing and magnitude of type I interferon responses by distinct sensors impact CD8 T cell exhaustion and chronic viral infection","volume":"11","author":"Wang","year":"2012","journal-title":"Cell Host Microbe"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/12\/6\/1150\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:54:10Z","timestamp":1760108050000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/12\/6\/1150"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,6,5]]},"references-count":53,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2024,6]]}},"alternative-id":["microorganisms12061150"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms12061150","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,6,5]]}}}