{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,20]],"date-time":"2025-11-20T12:56:03Z","timestamp":1763643363463,"version":"build-2065373602"},"reference-count":39,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2021,12,9]],"date-time":"2021-12-09T00:00:00Z","timestamp":1639008000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Central Public-interest Scientific Institution Basal Research Fund; Green Fish Drug Innovation Center Running Expense.","award":["CAFS (NO.2019A006);CAFS\uff08NO.2020TD11\uff09"],"award-info":[{"award-number":["CAFS (NO.2019A006);CAFS\uff08NO.2020TD11\uff09"]}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["NO.2017YFC1600704"],"award-info":[{"award-number":["NO.2017YFC1600704"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Lactic acid bacteria (LAB) are commonly applied to fish as a means of growth promotion and disease prevention. However, evidence regarding whether LAB colonize the gastrointestinal (GI) tract of fish remains sparse and controversial. Here, we investigated whether Lacticaseibacillus casei ATCC 393 (Lc) can colonize the GI tract of crucian carp. Sterile feed irradiated with 60Co was used to eliminate the influence of microbes, and 100% rearing water was renewed at 5-day intervals to reduce the fecal\u2013oral circulation of microbes. The experiment lasted 47 days and was divided into three stages: the baseline period (21 days), the administration period (7 days: day \u22126 to 0) and the post-administration period (day 1 to 19). Control groups were fed a sterile basal diet during the whole experimental period, whereas treatment groups were fed with a mixed diet containing Lc (1 \u00d7 107 cfu\/g) and spore of Geobacillus stearothermophilus (Gs, 1 \u00d7 107 cfu\/g) during the administration period and a sterile basal diet during the baseline and post-administration periods. An improved and highly sensitive selective culture method (SCM) was employed in combination with a transit marker (a Gs spore) to monitor the elimination of Lc in the GI tract. The results showed that Lc (&lt;2 cfu\/gastrointestine) could not be detected in any of the fish sampled from the treatment group 7 days after the cessation of the mixed diet, whereas Gs could still be detected in seven out of nine fish at day 11 and could not be detected at all at day 15. Therefore, the elimination speed of Lc was faster than that of the transit marker. Furthermore, high-throughput sequencing analysis combined with SCM was used to reconfirm the elimination kinetics of Lc in the GI tract. The results show that the Lc in the crucian carp GI tract, despite being retained at low relative abundance from day 7 (0.11% \u00b1 0.03%) to 21, was not viable. The experiments indicate that Lc ATCC 393 cannot colonize the GI tract of crucian carp, and the improved selective culture in combination with a transit marker represents a good method for studying LAB colonization of fish.<\/jats:p>","DOI":"10.3390\/microorganisms9122547","type":"journal-article","created":{"date-parts":[[2021,12,9]],"date-time":"2021-12-09T21:46:58Z","timestamp":1639086418000},"page":"2547","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Lacticaseibacillus casei ATCC 393 Cannot Colonize the Gastrointestinal Tract of Crucian Carp"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4929-871X","authenticated-orcid":false,"given":"Hongyu","family":"Zhang","sequence":"first","affiliation":[{"name":"Fishery Resource and Environment Research Center, Chinese Academy of Fishery Sciences, Beijing 100141, China"},{"name":"Chinese Academy of Fishery Sciences, Beijing 100141, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiyan","family":"Mu","sequence":"additional","affiliation":[{"name":"Fishery Resource and Environment Research Center, Chinese Academy of Fishery Sciences, Beijing 100141, China"},{"name":"Chinese Academy of Fishery Sciences, Beijing 100141, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongwei","family":"Wang","sequence":"additional","affiliation":[{"name":"Chinese Academy of Fishery Sciences, Beijing 100141, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haibo","family":"Wang","sequence":"additional","affiliation":[{"name":"Fishery Resource and Environment Research Center, Chinese Academy of Fishery Sciences, Beijing 100141, China"},{"name":"Chinese Academy of Fishery Sciences, Beijing 100141, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hui","family":"Wang","sequence":"additional","affiliation":[{"name":"Fishery Resource and Environment Research Center, Chinese Academy of Fishery Sciences, Beijing 100141, China"},{"name":"Chinese Academy of Fishery Sciences, Beijing 100141, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yingren","family":"Li","sequence":"additional","affiliation":[{"name":"Fishery Resource and Environment Research Center, Chinese Academy of Fishery Sciences, Beijing 100141, China"},{"name":"Chinese Academy of Fishery Sciences, Beijing 100141, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yingchun","family":"Mu","sequence":"additional","affiliation":[{"name":"Chinese Academy of Fishery Sciences, Beijing 100141, China"},{"name":"Key Laboratory of Control of Quality and Safety for Aquatic Products (Ministry of Agriculture and Rural Affairs), Chinese Academy of Fishery Sciences, Beijing 100141, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jinlong","family":"Song","sequence":"additional","affiliation":[{"name":"Chinese Academy of Fishery Sciences, Beijing 100141, China"},{"name":"Key Laboratory of Control of Quality and Safety for Aquatic Products (Ministry of Agriculture and Rural Affairs), Chinese Academy of Fishery Sciences, Beijing 100141, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lei","family":"Xia","sequence":"additional","affiliation":[{"name":"Chinese Academy of Fishery Sciences, Beijing 100141, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.fsi.2021.07.007","article-title":"The functionality of probiotics in aquaculture: An overview","volume":"117","author":"Alagawany","year":"2021","journal-title":"Fish Shellfish. Immun."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"737050","DOI":"10.1016\/j.aquaculture.2021.737050","article-title":"Therapeutic modulation of fish gut microbiota, a feasible strategy for aquaculture?","volume":"544","author":"Cicala","year":"2021","journal-title":"Aquacultrue"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1111\/raq.12248","article-title":"The adherence and colonization of microorganisms in fish gastrointestinal tract","volume":"11","author":"Li","year":"2019","journal-title":"Rev. Aquacult."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Tian, J., Kang, Y., Chu, G., Liu, H., Kong, Y., Zhao, L., Kong, Y., Shan, X., and Wang, G. (2020). Oral administration of lactobacillus casei expressing flagellin a protein confers effective protection against Aeromonas Veronii in common carp, Cyprinus Carpio. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21010033"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1007\/s12602-017-9297-3","article-title":"Combined or individual effects of dietary probiotic Pedicoccus acidilactici and nucleotide on growth performance, intestinal microbiota, hemato-biochemical parameters, and innate immune response in goldfish (Carassius auratus)","volume":"10","author":"Mehdinejad","year":"2018","journal-title":"Probiot. Antimicrob. Protein"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1007\/s12562-019-01385-1","article-title":"Adhesion and colonization properties of potentially probiotic Bacillus paralicheniformis strain FA6 isolated from grass carp intestine","volume":"86","author":"Zhao","year":"2020","journal-title":"Fisheries Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1007\/s12602-017-9373-8","article-title":"Changes in immunity, expression of some immune-related genes of shabot fish, Tor grypus, following experimental infection with Aeromonas hydrophila: Effects of autochthonous probiotics","volume":"10","author":"Mohammadian","year":"2018","journal-title":"Probiot. Antimicrob. Protein"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1823","DOI":"10.1016\/j.vaccine.2011.12.132","article-title":"Expression of infectious pancreatic necrosis virus (IPNV) VP2\u2013VP3 fusion protein in Lactobacillus casei and immunogenicity in rainbow trouts","volume":"30","author":"Zhao","year":"2012","journal-title":"Vaccine"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1111\/j.1574-695X.2007.00294.x","article-title":"Enhancement of the immune response and protection induced by probiotic lactic acid bacteria against furunculosis in rainbow trout (Oncorhynchus mykiss)","volume":"51","author":"Blas","year":"2007","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"13195","DOI":"10.1038\/s41598-017-13466-1","article-title":"Anti-infective effect of adhesive probiotic lactobacillus in fish is correlated with their spatial distribution in the intestinal tissue","volume":"7","author":"He","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.fsi.2018.11.022","article-title":"Effects of Lactococcus lactis subsp. lactis JCM5805 on colonization dynamics of gut microbiota and regulation of immunity in early ontogenetic stages of tilapia","volume":"86","author":"Xia","year":"2019","journal-title":"Fish Shellfish. Immun."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"714","DOI":"10.1016\/j.fsi.2019.02.003","article-title":"Spatiotemporal distribution of Streptococcus agalactiae attenuated vaccine strain YM001 in the intestinal tract of tilapia and its effect on mucosal associated immune cells","volume":"87","author":"Huang","year":"2019","journal-title":"Fish Shellfish. Immun."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1017\/S0007114507432986","article-title":"Changes in intestinal microbiota and humoral immune response following probiotic administration in brown trout (Salmo trutta)","volume":"97","author":"Balcazar","year":"2007","journal-title":"Br. J. Nutr."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3479","DOI":"10.1007\/s00253-014-6351-x","article-title":"Zebrafish gut colonization by mCherry-labelled lactic acid bacteria","volume":"99","author":"Russo","year":"2015","journal-title":"Appl. Microbiol. Biot."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/S1050-4648(03)00023-8","article-title":"Immune enhancement in rainbow trout (Oncorhynchus mykiss) by potential probiotic bacteria (Lactobacillus rhamnosus)","volume":"15","author":"Nikoskelainen","year":"2003","journal-title":"Fish Shellfish. Immun."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/S0044-8486(97)00299-8","article-title":"Lactic acid bacteria in fish: A review","volume":"160","author":"Gatesoupe","year":"1998","journal-title":"Aquaculture"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1111\/jam.14628","article-title":"Probiotics, lactic acid bacteria and bacilli: Interesting supplementation for aquaculture","volume":"129","author":"Lee","year":"2020","journal-title":"J. Appl. Microbiol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Conway, T., and Cohen, P.S. (2015). Commensal and pathogenic Escherichia coli metabolism in the gut. Microbiol. Spectr., 3.","DOI":"10.1128\/microbiolspec.MBP-0006-2014"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/S0044-8486(99)00187-8","article-title":"The use of probiotics in aquaculture","volume":"180","author":"Gatesoupe","year":"1999","journal-title":"Aquaculture"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1016\/S0969-806X(99)00400-4","article-title":"A study on radiation sterilization of SPF animal feed","volume":"57","author":"Chen","year":"2000","journal-title":"Radiat. Phys. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.12938\/bifidus1996.17.1","article-title":"Pharmacokinetics of probiotics and biotherapeutic agents in humans","volume":"17","author":"Marteau","year":"1998","journal-title":"Biosci. Microflora"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.mib.2018.10.005","article-title":"Colonization of the mammalian intestinal tract by enterococci","volume":"47","author":"Banla","year":"2019","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1016\/j.fsi.2017.06.017","article-title":"Apple cider vinegar boosted immunomodulatory and health promoting effects of Lactobacillus casei in common carp (Cyprinus carpio)","volume":"67","author":"Safari","year":"2017","journal-title":"Fish Shellfish. Immunol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1530\/REP-13-0141","article-title":"Comparison of fecundity and offspring immunity in zebrafish fed Lactobacillus rhamnosus CICC 6141 and Lactobacillus casei BL23","volume":"147","author":"Qin","year":"2013","journal-title":"Reproduction"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2771","DOI":"10.1128\/aem.61.7.2771-2774.1995","article-title":"Genetic marking of Lactococcus lactis shows its survival in the human gastrointestinal tract","volume":"61","author":"Klijn","year":"1995","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"823","DOI":"10.1046\/j.1365-2036.2000.00763.x","article-title":"Pharmacokinetics of Lactobacillus plantarum NCIMB 8826, Lactobacillus fermentum KLD, and Lactococcus lactis MG 1363 in the human gastrointestinal tract","volume":"14","author":"Vesa","year":"2000","journal-title":"Aliment. Pharmacol. Ther."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Zhang, H., Wang, H., Hu, K., Jiao, L., Zhao, M., Yang, X., and Xia, L. (2019). Effect of Dietary Supplementation of Lactobacillus Casei YYL3 and L. Plantarum YYL5 on Growth, Immune Response and Intestinal Microbiota in Channel Catfish. Animals, 9.","DOI":"10.3390\/ani9121005"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1111\/anu.12346","article-title":"Effect of dietary components on the gut microbiota of aquatic animals. A never-ending story?","volume":"22","author":"Zhou","year":"2016","journal-title":"Aquacult. Nutr."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1111\/j.1365-2095.2009.00689.x","article-title":"Probiotic applications for rainbow trout (Oncorhynchus mykiss Walbaum) I. Effects on growth performance, feed utilization, intestinal microbiota and related health criteria","volume":"16","author":"Merrifield","year":"2010","journal-title":"Aquacult. Nutr."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1111\/j.1365-2095.2009.00688.x","article-title":"Probiotic applications for rainbow trout (Oncorhynchus mykiss Walbaum) II. Effects on growth performance, feed utilization, intestinal microbiota and related health criteria postantibiotic treatment","volume":"16","author":"Merrifield","year":"2010","journal-title":"Aquacult. Nutr."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1315","DOI":"10.1016\/j.clnu.2019.05.014","article-title":"Mining Lactobacillus and Bifidobacterium for organisms with long-term gut colonization potential","volume":"39","author":"Xiao","year":"2020","journal-title":"Clin. Nutr."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1111\/j.1365-2036.2006.02749.x","article-title":"The effect of a probiotic drink with Lactobacillus plantarum 299v on the bacterial composition in faeces and mucosal biopsies of rectum and ascending colon","volume":"23","author":"Goossens","year":"2006","journal-title":"Aliment. Pharmacol. Ther."},{"key":"ref_33","unstructured":"De Vos, P., Garrity, G.M., Jones, D., Krieg, N.R., Ludwig, W., Rainey, F.A., Schleifer, K.-H., and Whitman, W.B. (2009). Bergey\u2019s Manual of Systematic Bacteriology, Springer."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1111\/jam.14961","article-title":"Spatial and temporal organization of jejunal microbiota in goats during animal development process","volume":"131","author":"Zhang","year":"2021","journal-title":"J. Appl. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Crobach, M.J.T., Ducarmon, Q.R., Terveer, E.M., Harmanus, C., Sanders, I.M.J.G., Verduin, K.M., Kuijper, E.J., and Zwittink, R.D. (2020). The bacterial gut microbiota of adult patients infected, colonized or noncolonized by clostridioides difficile. Microorganisms, 8.","DOI":"10.3390\/microorganisms8050677"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.jcrc.2018.04.013","article-title":"Comparison of traditional microbiological culture and 16S polymerase chain reaction analyses for identification of preoperative airway colonization for patients undergoing lung resection","volume":"46","author":"Howitt","year":"2018","journal-title":"J. Crit. Care"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.humic.2018.06.001","article-title":"Metagenomic and clinical microbiology","volume":"9","author":"Amrane","year":"2018","journal-title":"Hum. Microbiome J."},{"key":"ref_38","first-page":"1136","article-title":"Colonization of Bacillus licheniformis A1 in intestine of Ictalurus punctatus","volume":"26","author":"Zhang","year":"2019","journal-title":"J. Fish. Sci. China"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1388","DOI":"10.1016\/j.cell.2018.08.041","article-title":"Personalized gut mucosal colonization resistance to empiric probiotics is associated with unique host and microbiome features","volume":"174","author":"Zmora","year":"2018","journal-title":"Cell"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/12\/2547\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:44:20Z","timestamp":1760168660000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/12\/2547"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,9]]},"references-count":39,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["microorganisms9122547"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms9122547","relation":{},"ISSN":["2076-2607"],"issn-type":[{"type":"electronic","value":"2076-2607"}],"subject":[],"published":{"date-parts":[[2021,12,9]]}}}