{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T02:47:16Z","timestamp":1790909236029,"version":"4.1.0"},"update-to":[{"DOI":"10.1371\/journal.ppat.1012199","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2024,5,9]],"date-time":"2024-05-09T00:00:00Z","timestamp":1715212800000}}],"reference-count":54,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2024,4,29]],"date-time":"2024-04-29T00:00:00Z","timestamp":1714348800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"The Second Century Fund (C2F), Chulalongkorn University"},{"name":"The 90th Anniversary of Chulalongkorn University Fund"},{"name":"Thailand Science Research and Innovation Fund, Chulalongkorn University"},{"name":"Chulalongkorn University under Ratchadaphisek Somphot Endowment Fund to the Center of Excellence for Molecular Biology and Genomics of Shrimp"},{"name":"The Office of the Permanent Secretary, Ministry of Higher Education, Science, Research and Innovation (OPS MHESI), Thailand Science Research and Innovation (TSRI), and Chulalongkorn University","award":["RGNS 65-013"],"award-info":[{"award-number":["RGNS 65-013"]}]},{"name":"Chulalongkorn University, a grant for development of new faculty staff, Ratchadaphiseksomphot Fund","award":["DNS 65_033_23_003_1"],"award-info":[{"award-number":["DNS 65_033_23_003_1"]}]}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p>The microsporidian <jats:italic>Enterocytozoon hepatopenaei<\/jats:italic> (EHP) is a fungi-related, spore-forming parasite. EHP infection causes growth retardation and size variation in shrimp, resulting in severe economic losses. Studies on shrimp immune response have shown that several antimicrobial peptides (AMPs) were upregulated upon EHP infection. Among those highly upregulated AMPs is c-type lysozyme (<jats:italic>Lv<\/jats:italic>Lyz-c). However, the immune signaling pathway responsible for <jats:italic>Lv<\/jats:italic>Lyz-c production in shrimp as well as its function against the EHP infection are still poorly understood. Here, we characterized major shrimp immune signaling pathways and found that Toll and JAK\/STAT pathways were up-regulated upon EHP infection. Knocking down of a <jats:italic>Domeless<\/jats:italic> (<jats:italic>DOME<\/jats:italic>) receptor in the JAK\/STAT pathways resulted in a significant reduction of the <jats:italic>Lv<\/jats:italic>Lyz-c and the elevation of EHP copy number. We further elucidated the function of <jats:italic>Lv<\/jats:italic>Lyz-c by heterologously expressing a recombinant <jats:italic>Lv<\/jats:italic>Lyz-c (r<jats:italic>Lv<\/jats:italic>Lyz-c) in an <jats:italic>Escherichia coli<\/jats:italic>. r<jats:italic>Lv<\/jats:italic>Lyz-c exhibited antibacterial activity against several bacteria such as <jats:italic>Bacillus subtilis<\/jats:italic> and <jats:italic>Vibrio parahaemolyticus<\/jats:italic>. Interestingly, we found an antifungal activity of r<jats:italic>Lv<\/jats:italic>Lyz-c against <jats:italic>Candida albican<\/jats:italic>, which led us to further investigate the effects of r<jats:italic>Lv<\/jats:italic>Lyz-c on EHP spores. Incubation of the EHP spores with r<jats:italic>Lv<\/jats:italic>Lyz-c followed by a chitin staining showed that the signals were dramatically decreased in a dose-dependent manner, suggesting that r<jats:italic>Lv<\/jats:italic>Lyz-c possibly digest a chitin coat on the EHP spores. Transmission electron microscopy analysis revealed that an endospore layer, which is composed mainly of chitin, was digested by r<jats:italic>Lv<\/jats:italic>Lyz-c. Lastly, we observed that EHP spores that were treated with r<jats:italic>Lv<\/jats:italic>Lyz-c showed a significant reduction of the spore germination rate. We hypothesize that thinning of the endospore of EHP would result in altered permeability, hence affecting spore germination. This work provides insights into shrimp immune signaling pathways responsible for <jats:italic>Lv<\/jats:italic>Lyz-c production and its anti-EHP property. This knowledge will serve as important foundations for developing EHP control strategies.<\/jats:p>","DOI":"10.1371\/journal.ppat.1012199","type":"journal-article","created":{"date-parts":[[2024,4,29]],"date-time":"2024-04-29T17:40:51Z","timestamp":1714412451000},"page":"e1012199","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":25,"title":["Immune signaling of Litopenaeus vannamei c-type lysozyme and its role during microsporidian Enterocytozoon hepatopenaei (EHP) infection"],"prefix":"10.1371","volume":"20","author":[{"given":"Nutthapon","family":"Sangklai","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Premruethai","family":"Supungul","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2036-232X","authenticated-orcid":true,"given":"Pattana","family":"Jaroenlak","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anchalee","family":"Tassanakajon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,4,29]]},"reference":[{"key":"ppat.1012199.ref001","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1146\/annurev-resource-100518-093750","article-title":"Economics of aquaculture policy and regulation","volume":"11","author":"JL Anderson","year":"2019","journal-title":"Annual Review of Resource Economics"},{"issue":"2004","key":"ppat.1012199.ref002","first-page":"92","article-title":"Introductions and movement of Penaeus vannamei and Penaeus stylirostris in Asia and the Pacific.","volume":"10","author":"M Briggs","year":"2004","journal-title":"RAP publication"},{"issue":"1","key":"ppat.1012199.ref003","doi-asserted-by":"crossref","first-page":"45","DOI":"10.3354\/dao02621","article-title":"Determination of the infectious nature of the agent of acute hepatopancreatic necrosis syndrome affecting penaeid shrimp","volume":"105","author":"L Tran","year":"2013","journal-title":"Diseases of Aquatic Organisms"},{"key":"ppat.1012199.ref004","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1023\/A:1018580301578","article-title":"Major viral diseases of the black tiger prawn (Penaeus monodon) in Thailand.","volume":"13","author":"TW Flegel","year":"1997","journal-title":"World Journal of Microbiology and Biotechnology"},{"issue":"1","key":"ppat.1012199.ref005","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.jip.2009.06.004","article-title":"Enterocytozoon hepatopenaei sp. nov.(Microsporida: Enterocytozoonidae), a parasite of the black tiger shrimp Penaeus monodon (Decapoda: Penaeidae): Fine structure and phylogenetic relationships.","volume":"102","author":"S Tourtip","year":"2009","journal-title":"Journal of Invertebrate Pathology"},{"issue":"2","key":"ppat.1012199.ref006","doi-asserted-by":"crossref","first-page":"85","DOI":"10.3354\/dao040085","article-title":"The haemocytic origin of lymphoid organ spheroid cells in the penaeid prawn Penaeus monodon","volume":"40","author":"MS Anggraeni","year":"2000","journal-title":"Diseases of Aquatic Organisms"},{"key":"ppat.1012199.ref007","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1746-6148-9-139","article-title":"The microsporidian Enterocytozoon hepatopenaei is not the cause of white feces syndrome in whiteleg shrimp Penaeus (Litopenaeus) vannamei.","volume":"9","author":"A Tangprasittipap","year":"2013","journal-title":"BMC veterinary research"},{"key":"ppat.1012199.ref008","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.aquaculture.2015.10.028","article-title":"Review of current disease threats for cultivated penaeid shrimp in Asia","volume":"452","author":"S Thitamadee","year":"2016","journal-title":"Aquaculture"},{"key":"ppat.1012199.ref009","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s12917-016-0923-1","article-title":"Laboratory cohabitation challenge model for shrimp hepatopancreatic microsporidiosis (HPM) caused by Enterocytozoon hepatopenaei (EHP).","volume":"13","author":"PV Salachan","year":"2016","journal-title":"BMC Veterinary Research"},{"key":"ppat.1012199.ref010","doi-asserted-by":"crossref","first-page":"739322","DOI":"10.1016\/j.aquaculture.2023.739322","article-title":"5-Aminolaevulinic acid reduced the mortality of the Pacific 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Destoumieux","year":"2000","journal-title":"Journal of Cell Science"},{"key":"ppat.1012199.ref038","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1016\/j.fsi.2019.01.051","article-title":"Proteomic and metabolomic responses in hepatopancreas of whiteleg shrimp Litopenaeus vannamei infected by microsporidian Enterocytozoon hepatopenaei","volume":"87","author":"M Ning","year":"2019","journal-title":"Fish & Shellfish Immunology"},{"key":"ppat.1012199.ref039","doi-asserted-by":"crossref","first-page":"108605","DOI":"10.1016\/j.fsi.2023.108605","article-title":"Transcriptome analysis of the hepatopancreas in Penaeus vannamei under experimental infection with Enterocytozoon hepatopenaei (EHP).","volume":"134","author":"Z Cao","year":"2023","journal-title":"Fish & Shellfish Immunology"},{"issue":"3","key":"ppat.1012199.ref040","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1111\/j.1365-2567.2005.02091.x","article-title":"Signal transducer and activator of transcription 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monodon.","volume":"13","author":"P Laohawutthichai","year":"2023","journal-title":"Scientific ReportsJun 17"},{"issue":"1","key":"ppat.1012199.ref050","doi-asserted-by":"crossref","first-page":"996","DOI":"10.1038\/s41598-023-27906-8","article-title":"White spot syndrome virus impact on the expression of immune genes and gut microbiome of black tiger shrimp Penaeus monodon","volume":"13","author":"T Jatuyosporn","year":"2023","journal-title":"Scientific Reports"},{"issue":"3","key":"ppat.1012199.ref051","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1016\/j.fsi.2005.06.005","article-title":"White shrimp (Litopenaeus vannamei) recombinant lysozyme has antibacterial activity against Gram negative bacteria: Vibrio alginolyticus, Vibrio parahemolyticus and Vibrio cholerae","volume":"20","author":"E De-La-Re-Vega","year":"2006","journal-title":"Fish & Shellfish 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