{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T23:36:09Z","timestamp":1792020969785,"version":"4.4.0"},"reference-count":21,"publisher":"Public Library of Science (PLoS)","issue":"1","license":[{"start":{"date-parts":[[2026,1,7]],"date-time":"2026-01-07T00:00:00Z","timestamp":1767744000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec001\">\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Cryopreservation and lyophilization are standard methods for preserving microbial cultures, but they are expensive and labor-intensive, creating a barrier for resource-constrained laboratories. This study evaluated the efficacy of a simple and inexpensive alternative: long-term storage in washed and sterilized sand.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Fourteen bacterial strains from ten genera were preserved in sealed tubes containing washed and sterilized sand at room temperature. Viability, assessed by colony-forming units (CFU\/g), was quantified at inoculation and after 7 and 14 years. Phenotypic stability was evaluated by comparing the pre- and post-storage profiles for enzyme activity and stress tolerance.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      After seven years, all 14 strains remained culturable with preserved phenotypic features. After 14 years, 13 strains (93%) were viable. The highest recovery rates were observed in spore-forming Gram-positive bacteria (e.g.,\n                      <jats:italic>Bacillus sp<\/jats:italic>\n                      .), indicating taxon-specific survival. Notably, the Gram-negative strain\n                      <jats:italic>Enterobacter hormaechei<\/jats:italic>\n                      remained stable with only a approximate 1.58 log\u2081\u2080 reduction over 14 years (half-life\u2009=\u20092.67 years). All resuscitated strains maintained their pre-storage characteristics. The distinct survival patterns for each species indicate that contamination was not the primary driver of the outcomes.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec004\">\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Washed and sterile sand offers a practical, low-energy matrix for long-term preservation of a diverse range of bacteria, effectively maintaining cultivability and key phenotypic attributes for over a decade. This approach is an economical preservation strategy for resource-limited laboratories and field collections.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pone.0340501","type":"journal-article","created":{"date-parts":[[2026,1,7]],"date-time":"2026-01-07T18:46:56Z","timestamp":1767811616000},"page":"e0340501","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["Long-term preservation of bacteria in washed sterile sand: A 14-year 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