{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T08:10:39Z","timestamp":1771920639150,"version":"3.50.1"},"reference-count":32,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T00:00:00Z","timestamp":1768262400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Encephalitozoon cuniculi<\/jats:italic>\n                      is an opportunistic pathogen with significant zoonotic potential, particularly for immunocompromised individuals. However, evidence-based disinfection protocols against its environmentally resistant spores are lacking, leading to potential reliance on suboptimal agents.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>\n                      This study aimed to establish a rigorous, data-driven hierarchy for the efficacy of common chemical disinfectants against\n                      <jats:italic>E. cuniculi<\/jats:italic>\n                      spores by integrating assessments of structural integrity, cellular infectivity, and\n                      <jats:italic>in vivo<\/jats:italic>\n                      dissemination.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We employed a multi-modal approach. Spores were treated with rapid-acting (75% ethanol, 1% hydrogen peroxide, chlorine-based agent) and long-acting (nano-silver, quaternary ammonium compounds\u2014QACs) disinfectants. Sporicidal effects were evaluated via flow cytometry using ethidium bromide (EB) staining. Functional infectivity was quantified in Vero cells using a species-specific TaqMan qPCR assay targeting intracellular spore DNA. A murine model was used to assess the capacity of treated spores to establish systemic infection, quantified by spore DNA load in blood and kidney tissue.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Flow cytometry revealed that 75% ethanol caused significant membrane damage (~51% EB-positive spores in 20\u202fmin) but failed to consistently abolish infectivity in cell culture or mice. Chlorine-based agents and hydrogen peroxide demonstrated potent, time-dependent inactivation, achieving &amp;gt;96 and 100% infectivity reduction in cells and mice, respectively, within 60\u202fmin. Notably, chlorine agents rendered spores non-infectious without inducing proportional EB uptake, indicating a mechanism distinct from gross membrane disruption. The long-acting disinfectant QACs necessitated an extended contact time (up to 5\u202fh) to achieve complete prevention of\n                      <jats:italic>in vivo<\/jats:italic>\n                      infection. This prolonged exposure induced substantial spore aggregation, thereby impeding accurate flow cytometric analysis.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      The chlorine-based disinfectant and hydrogen peroxide (rapid-acting), as well as the quaternary ammonium compound (long-acting), all effectively eliminated the infectivity of\n                      <jats:italic>E. cuniculi<\/jats:italic>\n                      spores. Crucially, we demonstrate a critical dissociation between spore structural damage and viability, highlighting the necessity of functional infectivity assays over structural metrics alone. This study provides a foundational framework for evidence-based infection control guidelines in clinical, veterinary, and public health settings.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmicb.2025.1744472","type":"journal-article","created":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T10:13:58Z","timestamp":1768299238000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Chemical disinfection of Encephalitozoon cuniculi: toward evidence-based infection control guidelines"],"prefix":"10.3389","volume":"16","author":[{"given":"Jianhua","family":"Gao","sequence":"first","affiliation":[{"name":"State Key Laboratory of Resource Insects, Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University","place":["Chongqing, China"]},{"name":"School of Medical Technology, Jiangxi Medical College","place":["Shangrao, Jiangxi, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xianzhi","family":"Meng","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Resource Insects, Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University","place":["Chongqing, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhangshuai","family":"He","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Resource Insects, Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University","place":["Chongqing, China"]},{"name":"College of Clinical Medicine, Chongqing Three Gorges Medical College","place":["Chongqing, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yunlin","family":"Tang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Resource Insects, Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University","place":["Chongqing, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jie","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Resource Insects, Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University","place":["Chongqing, China"]},{"name":"College of Sericulture, Textile and Biomass Sciences, Southwest University","place":["Chongqing, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xuemei","family":"He","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Resource Insects, Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University","place":["Chongqing, China"]},{"name":"College of Clinical Medicine, Chongqing Three Gorges Medical College","place":["Chongqing, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hua","family":"Cao","sequence":"additional","affiliation":[{"name":"School of Medical Technology, Jiangxi Medical College","place":["Shangrao, Jiangxi, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuyan","family":"Long","sequence":"additional","affiliation":[{"name":"School of Medical Technology, Jiangxi Medical College","place":["Shangrao, Jiangxi, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guoqing","family":"Pan","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Resource Insects, Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University","place":["Chongqing, China"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2026,1,13]]},"reference":[{"key":"ref1","first-page":"1977","article-title":"Weakening effect of cell permeabilizers on gram-negative bacteria causing biodeterioration","volume":"66","author":"Alakomi","year":"2000","journal-title":"Appl. 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