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Several phage-based products have been approved for agri-food applications, all of which rely on strictly lytic phages to ensure safety, regulatory compliance, and rapid bactericidal action. Despite its industrial relevance, only two phages targeting the genus\n                      <jats:italic toggle=\"yes\">Alicyclobacillus<\/jats:italic>\n                      , thermoacidophilic spore-formers responsible for fruit product spoilage, have been reported to date, leaving a very limited arsenal for their control. This study reports the\n                      <jats:italic toggle=\"yes\">Alicyclobacillus<\/jats:italic>\n                      phage MMB025, infecting\n                      <jats:italic toggle=\"yes\">Alicyclobacillus acidoterrestris<\/jats:italic>\n                      , a species associated with spoilage. Transmission electron microscopy revealed a contractile-tailed morphotype, and host range assays demonstrated specificity for a narrow subset of\n                      <jats:italic toggle=\"yes\">Alicyclobacillus<\/jats:italic>\n                      isolates. MMB025 displayed stability across a wide pH range (3\u201312) and moderate heat (\u226460\u00b0C). Hybrid sequencing identified a 105 kb dsDNA genome encoding structural, replication, and lysogeny modules, the latter indicating a temperate lifestyle. Consistently, the phage genome showed 99.99% identity to a chromosomal region of\n                      <jats:italic toggle=\"yes\">A. acidoterrestris<\/jats:italic>\n                      DSM 3922\n                      <jats:sup>T<\/jats:sup>\n                      , most likely corresponding to MMB025 prophage state. Genome mapping showed prophage integration within the sporulation sigma factor gene\n                      <jats:italic toggle=\"yes\">sigK<\/jats:italic>\n                      , reminiscent of the\n                      <jats:italic toggle=\"yes\">Bacillus subtilis skin<\/jats:italic>\n                      element, suggesting a novel phage-sporulation interaction in\n                      <jats:italic toggle=\"yes\">Alicyclobacillus<\/jats:italic>\n                      . Despite the presence of lysogeny-related features, bioinformatic lifestyle predictions were inconclusive, emphasizing current limitations of\n                      <jats:italic toggle=\"yes\">in silico<\/jats:italic>\n                      classification. Although temperate phages like MMB025 are not ideal for direct biocontrol, the genome encodes putative lysins and enzymes with potential for spoilage mitigation. This study contributes to expanding the still-limited diversity of\n                      <jats:italic toggle=\"yes\">Alicyclobacillus<\/jats:italic>\n                      phages and provides new insights into phage-host interactions in thermoacidophilic spore-forming bacteria.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      The genus\n                      <jats:italic toggle=\"yes\">Alicyclobacillus<\/jats:italic>\n                      remains a persistent problem for the fruit juice industry, as its thermoacidophilic nature leads to spoilage events that cause product recalls, loss of consumer confidence, and significant economic losses. Yet remarkably little is known about the phages infecting these bacteria, with only two described to date. This study reports the first infectious phage with genomic identity to a prophage in the genus, the\n                      <jats:italic toggle=\"yes\">Alicyclobacillus<\/jats:italic>\n                      phage MMB025. This finding broadens the extremely limited record of\n                      <jats:italic toggle=\"yes\">Alicyclobacillus<\/jats:italic>\n                      phages and reveals integration within the sporulation sigma factor\n                      <jats:italic toggle=\"yes\">sigK<\/jats:italic>\n                      , suggesting a novel link between prophage activity and sporulation regulation in\n                      <jats:italic toggle=\"yes\">Alicyclobacillus<\/jats:italic>\n                      . By combining stability assays, host range testing, and genome analysis, MMB025 represents both a valuable model for phage-spore-former biology and a genetic resource for candidate lysins with potential applications in food preservation.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.03747-25","type":"journal-article","created":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T13:00:38Z","timestamp":1775221238000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["A novel temperate phage from\n                    <i>Alicyclobacillus<\/i>\n                    : first evidence in this genus of genomic identity to a\n                    <i>sigK<\/i>\n                    -integrated prophage"],"prefix":"10.1128","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8516-2487","authenticated-orcid":false,"given":"In\u00eas 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