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Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      Food contamination by aflatoxin-producing\n                      <jats:italic>Aspergillus<\/jats:italic>\n                      spp. remains a major food safety and public health concern.\n                      <jats:italic>Trichoderma<\/jats:italic>\n                      spp. and their metabolites are promising biocontrol agents, yet the volatile organic compound (VOC)-mediated interactions underlying growth inhibition of\n                      <jats:italic>Aspergillus<\/jats:italic>\n                      spp. are not fully understood.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We applied untargeted metabolomics using Headspace Solid Phase Microextraction Gas Chromatography\u2013Mass Spectrometry (HS-SPME-GC-MS) and multivariate analyses to profile VOCs from three\n                      <jats:italic>Trichoderma<\/jats:italic>\n                      strains (\n                      <jats:italic>T. asperellum<\/jats:italic>\n                      BTU,\n                      <jats:italic>T. virens<\/jats:italic>\n                      OSK13,\n                      <jats:italic>T. harzianum<\/jats:italic>\n                      OSK34) in monoculture and in co-culture with\n                      <jats:italic>Aspergillus flavus<\/jats:italic>\n                      NRRL 3357 and\n                      <jats:italic>A. parasiticus<\/jats:italic>\n                      B62. Principal component analysis (PCA), ANOVA, PLS-DA, and Spearman correlation were used to characterize VOC profiles and identify discriminant compounds.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Co-cultures produced 30 VOCs across 9\u201312 chemical classes, dominated by sesquiterpenes, alcohols, and ketones. Among the\n                      <jats:italic>Trichoderma<\/jats:italic>\n                      strains, BTU (against\n                      <jats:italic>A. flavus<\/jats:italic>\n                      3357) and OSK34 (against\n                      <jats:italic>A. parasiticus<\/jats:italic>\n                      B62) showed the greatest VOC abundance and diversity. PCA confirmed\n                      <jats:italic>Trichoderma<\/jats:italic>\n                      -driven changes in VOC profiles during co-cultures, with OSK13 and OSK34 sharing greater VOC profile similarity compared to BTU. In addition,\n                      <jats:italic>Trichoderma<\/jats:italic>\n                      spp. produced both species-specific and shared VOCs, with some also shared with\n                      <jats:italic>Aspergillus<\/jats:italic>\n                      spp. ANOVA further identified significant upregulation and downregulation of VOCs during co-culture. Finally, PLS-DA, ANOVA, and Spearman correlation identified four key discriminant VOCs: 6-pentyl-2H-pyran-2-one, 2-pentylfuran, 1-decanol, and 1-ethenyl-4-methoxybenzene. These VOCs were most abundant in BTU-\n                      <jats:italic>Aspergillus<\/jats:italic>\n                      co-cultures with observed antifungal activity.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      These findings provide insight into potential mechanisms of\n                      <jats:italic>Aspergillus<\/jats:italic>\n                      growth inhibition by\n                      <jats:italic>Trichoderma<\/jats:italic>\n                      spp. through comparison of monocultures to co-cultures. In addition, the four identified key discriminant VOCs make promising VOC targets for selecting\n                      <jats:italic>Trichoderma<\/jats:italic>\n                      strains for biocontrol of toxigenic\n                      <jats:italic>Aspergillus<\/jats:italic>\n                      , particularly\n                      <jats:italic>A. flavus<\/jats:italic>\n                      . Together, this study establishes a metabolomic framework for VOC-mediated biocontrol and a basis for optimizing\n                      <jats:italic>Trichoderma<\/jats:italic>\n                      selection for biocontrol purposes. Future research should validate the bioactivity roles of these key VOCs and their effect on aflatoxin production, test their antifungal mechanisms, examine synergistic effects, and assess environmental influences to strengthen metabolomics-informed biocontrol strategies for reducing aflatoxin contamination in food systems.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmicb.2026.1873321","type":"journal-article","created":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T05:30:00Z","timestamp":1791523800000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Signaling in the gas phase: comparative volatile profiling of biocontrol Trichoderma spp. and aflatoxin-producing Aspergillus spp. in isolation and co-culture using HS-SPME GC-MS"],"prefix":"10.3389","volume":"17","author":[{"given":"Zilfa","family":"Irakoze","sequence":"first","affiliation":[{"name":"Department of Food Science, The Pennsylvania State University","place":["University Park, PA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nataliya M.","family":"Voloshchuk","sequence":"additional","affiliation":[{"name":"Department of Food Science, The Pennsylvania State University","place":["University Park, PA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joshua D.","family":"Lambert","sequence":"additional","affiliation":[{"name":"Department of Food Science, The Pennsylvania State University","place":["University Park, PA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helene","family":"Hopfer","sequence":"additional","affiliation":[{"name":"Department of Food Science, The Pennsylvania State University","place":["University Park, PA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Josephine","family":"Wee","sequence":"additional","affiliation":[{"name":"Department of Food Science, The Pennsylvania State University","place":["University Park, PA, United States"]},{"name":"One Health Microbiome Center, Huck Institutes of the Life Sciences, The Pennsylvania State University","place":["University Park, PA, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,10,9]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"2069","DOI":"10.3389\/fmicb.2019.02069","article-title":"Potential of Atoxigenic Aspergillus flavus vegetative compatibility groups associated with maize and groundnut in ghana as biocontrol agents for Aflatoxin management.","volume":"10","author":"Agbetiameh","year":"2019","journal-title":"Front. 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