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Bacteriophage viruses (phages) offer renewed therapeutic promise, yet their performance in\u00a0vivo often differs from expectations based on standard laboratory assays. Biofilms, with their dense extracellular matrix and spatial heterogeneity, restrict phage access, diffusion, and replication. Anaerobic niches and metabolically dormant bacterial subpopulations at the biofilm core pose additional hurdles, often rendering bacteria tolerant to both antibiotics and phages. These physiological constraints are rarely captured during phage isolation, where agar\u2010based plaque assays, oxygen\u2010rich conditions, and rapidly growing hosts inadvertently deselect phages adapted to biofilms, anaerobic zones, or dormant states. Understanding how phage\u2013bacterial interactions vary across these niches and how antibiotics synergize with or antagonize phage activity is essential for designing therapies with improved effectiveness beyond the Petri dish. This TechNote synthesizes key challenges for phage infection dynamics in biofilms, anaerobic environments, and dormant populations, highlighting the need for isolation and screening strategies that better reflect the physiology of clinical infections.<\/jats:p>","DOI":"10.1111\/apm.70249","type":"journal-article","created":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T02:32:37Z","timestamp":1787711557000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["<scp>PMI TechNote<\/scp>\n                    : Bacteriophages\u2014From the Petri Dish to Conquering the Nooks and Crannies of Difficult\u2010to\u2010Treat Infections"],"prefix":"10.1111","volume":"134","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-3673-4737","authenticated-orcid":false,"given":"Amalie H\u00f8gh","family":"Eichler","sequence":"first","affiliation":[{"name":"Department of Immunology and Microbiology University of Copenhagen  Copenhagen Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2699-9858","authenticated-orcid":false,"given":"Ida Friberg","family":"Hitz","sequence":"additional","affiliation":[{"name":"Department of Plant and Environmental Sciences University of Copenhagen  Frederiksberg C Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7025-8558","authenticated-orcid":false,"given":"Kira C\u00e9line","family":"Koonce","sequence":"additional","affiliation":[{"name":"Department of Plant and Environmental Sciences University of Copenhagen  Frederiksberg C Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8223-8407","authenticated-orcid":false,"given":"Leah M.","family":"Smith","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology University of Otago  Dunedin New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3711-914X","authenticated-orcid":false,"given":"Jesper Juel","family":"Mauritzen","sequence":"additional","affiliation":[{"name":"Department of Immunology and Microbiology University of Copenhagen  Copenhagen Denmark"},{"name":"Department of Plant and Environmental Sciences University of Copenhagen  Frederiksberg C Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5904-8859","authenticated-orcid":false,"given":"Nina Molin","family":"H\u00f8yland\u2010Kroghsbo","sequence":"additional","affiliation":[{"name":"Department of Immunology and Microbiology University of Copenhagen  Copenhagen Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,8,25]]},"reference":[{"key":"e_1_2_15_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140\u20106736(24)01867\u20101"},{"key":"e_1_2_15_3_1","unstructured":"Antimicrobial Resistance Division (AMR)andWHO \u201cGlobal Antibiotic Resistance Surveillance Report 2025 WHO Global Antimicrobial Resistance and Use Surveillance System (GLASS) \u201d(2025) 1\u2013114."},{"key":"e_1_2_15_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro3564"},{"key":"e_1_2_15_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1473\u20103099(21)00612\u20105"},{"key":"e_1_2_15_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/NRMICRO2315"},{"key":"e_1_2_15_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/S41564\u2010024\u201001705\u2010X"},{"key":"e_1_2_15_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2026.117014"},{"key":"e_1_2_15_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2020.03.016"},{"key":"e_1_2_15_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/J.CHOM.2015.10.013"},{"key":"e_1_2_15_11_1","volume-title":"Bacteriophages: Methods and Protocols","author":"Clokie M. 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