{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,30]],"date-time":"2026-10-30T15:50:16Z","timestamp":1793375416896,"version":"4.5.0"},"reference-count":47,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,1,19]],"date-time":"2022-01-19T00:00:00Z","timestamp":1642550400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>In this study, we examined the effect of a bacteriophage cocktail (tentatively designated as the Foodborne Outbreak Pill (FOP)) on the levels of Listeria monocytogenes in simulated small intestine, large intestine, and Caco-2 model systems. We found that FOP survival during simulated passage of the upper gastrointestinal was dependent on stomach pH, and that FOP robustly inhibited L. monocytogenes levels with effectiveness comparable to antibiotic treatment (ampicillin) under simulated ilium and colon conditions. The FOP did not inhibit the commensal bacteria, whereas ampicillin treatment led to dysbiosis-like conditions. The FOP was also more effective than an antibiotic in protecting Caco-2 cells from adhesion and invasion by L. monocytogenes (5-log reduction vs. 1-log reduction) while not triggering an inflammatory response. Our data suggested that the FOP may provide a robust protection against L. monocytogenes should the bacterium enter the human gastrointestinal tract (e.g., by consumption of contaminated food), without deleterious impact on the commensal bacteria.<\/jats:p>","DOI":"10.3390\/v14020190","type":"journal-article","created":{"date-parts":[[2022,1,19]],"date-time":"2022-01-19T21:02:27Z","timestamp":1642626147000},"page":"190","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["A Bacteriophage Cocktail Significantly Reduces Listeria monocytogenes without Deleterious Impact on the Commensal Gut Microbiota under Simulated Gastrointestinal Conditions"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7416-1358","authenticated-orcid":false,"given":"Rasmus Riemer","family":"Jakobsen","sequence":"first","affiliation":[{"name":"Section of Microbiology and Fermentation, Department of Food Science, Faculty of Science, University of Copenhagen, 1958 Frederiksberg, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jimmy T.","family":"Trinh","sequence":"additional","affiliation":[{"name":"Intralytix, Inc., 8681 Robert Fulton Drive, Columbia, MD 21046, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Louise","family":"Bomholtz","sequence":"additional","affiliation":[{"name":"Section of Microbiology and Fermentation, Department of Food Science, Faculty of Science, University of Copenhagen, 1958 Frederiksberg, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Signe Kristine","family":"Brok-Lauridsen","sequence":"additional","affiliation":[{"name":"Section of Microbiology and Fermentation, Department of Food Science, Faculty of Science, University of Copenhagen, 1958 Frederiksberg, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Sulakvelidze","sequence":"additional","affiliation":[{"name":"Intralytix, Inc., 8681 Robert Fulton Drive, Columbia, MD 21046, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8121-1114","authenticated-orcid":false,"given":"Dennis Sandris","family":"Nielsen","sequence":"additional","affiliation":[{"name":"Section of Microbiology and Fermentation, Department of Food Science, Faculty of Science, University of Copenhagen, 1958 Frederiksberg, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,19]]},"reference":[{"key":"ref_1","unstructured":"ECDC (2020). Listeriosis, Annual Epidemiological Report for 2017."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1073","DOI":"10.1016\/S1473-3099(14)70870-9","article-title":"The global burden of listeriosis: A systematic review and meta-analysis","volume":"14","author":"Devleesschauwer","year":"2014","journal-title":"Lancet Infect. Dis."},{"key":"ref_3","first-page":"694","article-title":"Experimental Listeria enteritis. I. An electron microscopic study of the epithelial phase in experimental listeria infection","volume":"26","author":"Tenner","year":"1972","journal-title":"Lab. Investig."},{"key":"ref_4","first-page":"391","article-title":"A bacteriophage cocktail targeting Escherichia coli reduces E. coli in simulated gut conditions, while preserving a non-targeted representative commensal normal microbiota","volume":"9","author":"Cieplak","year":"2018","journal-title":"Gut Microbes"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Domingo-Calap, P., and Delgado-Mart\u00ednez, J. (2018). Bacteriophages: Protagonists of a post-antibiotic era. Antibiotics, 7.","DOI":"10.3390\/antibiotics7030066"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.mib.2019.03.005","article-title":"Stronger together? Perspectives on phage-antibiotic synergy in clinical applications of phage therapy","volume":"51","author":"Segall","year":"2019","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/j.copbio.2010.10.008","article-title":"Bacteriophages as biocontrol agents of food pathogens","volume":"22","author":"Mahony","year":"2011","journal-title":"Curr. Opin. Biotechnol."},{"key":"ref_8","unstructured":"United States Food and Drug Administration Center for Food Safety and Applied Nutrition (2021, December 05). Agency Response Letter, GRAS Notice No. 000198, Available online: http:\/\/www.cfsan.fda.gov\/~rdb\/opa-g198.html."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.cofs.2020.10.006","article-title":"The use of bacteriophages for food safety","volume":"36","author":"Endersen","year":"2020","journal-title":"Curr. Opin. Food Sci."},{"key":"ref_10","first-page":"358051","article-title":"What is new in Listeriosis?","volume":"2014","year":"2014","journal-title":"BioMed Res. Int."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/j.chom.2019.01.014","article-title":"Phage Therapy: A Renewed Approach to Combat Antibiotic-Resistant Bacteria","volume":"25","author":"Kortright","year":"2019","journal-title":"Cell Host Microbe"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2000","DOI":"10.1002\/med.21572","article-title":"Phage therapy: What factors shape phage pharmacokinetics and bioavailability? Systematic and critical review","volume":"39","year":"2019","journal-title":"Med. Res. Rev."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1336","DOI":"10.4315\/0362-028X.JFP-18-587","article-title":"A Bacteriophage Cocktail Eliminates Salmonella Typhimurium from the Human Colonic Microbiome while Preserving Cytokine Signaling and Preventing Attachment to and Invasion of Human Cells by Salmonella In Vitro","volume":"82","author":"Moye","year":"2019","journal-title":"J. Food Prot."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e00497-21","DOI":"10.1128\/Spectrum.00497-21","article-title":"Prophylactic Administration of a Bacteriophage Cocktail Is Safe and Effective in Reducing Salmonella enterica Serovar Typhimurium Burden in Vivo","volume":"9","author":"Chaffringeon","year":"2021","journal-title":"Microbiol. Spectr."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1101\/SQB.1953.018.01.039","article-title":"Some problems of animal virology as studied by the plaque technique","volume":"18","author":"Dulbecco","year":"1953","journal-title":"Cold Spring Harb. Symp. Quant. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"fny231","DOI":"10.1093\/femsle\/fny231","article-title":"The Smallest Intestine (TSI)-a low volume in vitro model of the small intestine with increased throughput","volume":"365","author":"Cieplak","year":"2018","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"8508","DOI":"10.1038\/srep08508","article-title":"No difference in small bowel microbiota between patients with irritable bowel syndrome and healthy controls","volume":"5","author":"Dlugosz","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"410","DOI":"10.3109\/00365521.2015.1116107","article-title":"Differences of microbiota in small bowel and faeces between irritable bowel syndrome patients and healthy subjects","volume":"51","author":"Chung","year":"2016","journal-title":"Scand. J. Gastroenterol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1111\/j.1472-765X.1991.tb00492.x","article-title":"Selection, differentiation and counting of haemolytic Listeria spp. on PALCAM medium","volume":"12","author":"Mossel","year":"1991","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e4268","DOI":"10.7717\/peerj.4268","article-title":"CoMiniGut\u2014A small volume in vitro colon model for the screening of gut microbial fermentation processes","volume":"6","author":"Wiese","year":"2018","journal-title":"PeerJ"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.mimet.2017.10.015","article-title":"Have you tried spermine? A rapid and cost-effective method to eliminate dextran sodium sulfate inhibition of PCR and RT-PCR","volume":"144","author":"Krych","year":"2018","journal-title":"J. Microbiol. Methods"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2460","DOI":"10.1093\/bioinformatics\/btq461","article-title":"Search and clustering orders of magnitude faster than BLAST","volume":"26","author":"Edgar","year":"2010","journal-title":"Bioinformatics"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Edgar, R.C. (2016). UNOISE2: Improved error-correction for Illumina 16S and ITS amplicon sequencing. bioRxiv, 081257.","DOI":"10.1101\/081257"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Edgar, R. (2016). SINTAX: A simple non-Bayesian taxonomy classifier for 16S and ITS sequences. bioRxiv, 074161.","DOI":"10.1101\/074161"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"716","DOI":"10.1099\/ijs.0.038075-0","article-title":"Introducing EzTaxon-e: A prokaryotic 16s rRNA gene sequence database with phylotypes that represent uncultured species","volume":"62","author":"Kim","year":"2012","journal-title":"Int. J. Syst. Evol. Microbiol."},{"key":"ref_26","unstructured":"Team RC (2019). R: A Language and Environment for Statistical Title. 4.0.1, R Foundation for Statistical Computing."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1200","DOI":"10.1038\/nmeth.2658","article-title":"Differential abundance analysis for microbial marker-gene surveys","volume":"10","author":"Paulson","year":"2013","journal-title":"Nat. Methods"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1186\/s40168-017-0237-y","article-title":"Normalization and microbial differential abundance strategies depend upon data characteristics","volume":"5","author":"Weiss","year":"2017","journal-title":"Microbiome"},{"key":"ref_29","first-page":"191","article-title":"metagenomeSeq: Statistical analysis for sparse high-throughput sequencing","volume":"1","author":"Paulson","year":"2014","journal-title":"Bioconductor Package"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"McMurdie, P.J., and Holmes, S. (2013). Phyloseq: An R Package for Reproducible Interactive Analysis and Graphics of Microbiome Census Data. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0061217"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1111\/j.1654-1103.2003.tb02228.x","article-title":"VEGAN, a package of R functions for community ecology","volume":"14","author":"Dixon","year":"2003","journal-title":"J. Veg. Sci."},{"key":"ref_32","unstructured":"Kassambara, A. (2021, December 05). \u201cggplot2\u201d Based Publication Ready Plots [R Package Ggpubr VERSION 0.4.0]. Available online: https:\/\/rpkgs.datanovia.com\/ggpubr\/."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1002\/wics.147","article-title":"ggplot2","volume":"3","author":"Wickham","year":"2011","journal-title":"Wiley Interdiscip. Rev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"7528","DOI":"10.1128\/AEM.71.11.7528-7530.2005","article-title":"Application of measurements of transepithelial electrical resistance of intestinal epithelial cell monolayers to evaluate probiotic activity","volume":"71","author":"Klingberg","year":"2005","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"5706","DOI":"10.1073\/pnas.1523899113","article-title":"Bacteriocin from epidemic Listeria strains alters the host intestinal microbiota to favor infection","volume":"113","author":"Quereda","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Becattini, S., and Pamer, E.G. (2018). Multifaceted defense against listeria monocytogenes in the gastro-intestinal lumen. Pathogens, 7.","DOI":"10.3390\/pathogens7010001"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"624234","DOI":"10.1155\/2010\/624234","article-title":"Bacteriophage administration reduces the concentration of Listeria monocytogenes in the gastrointestinal tract and its translocation to spleen and liver in experimentally infectedmice","volume":"2010","author":"Mai","year":"2010","journal-title":"Int. J. Microbiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1093\/femsre\/fuaa020","article-title":"Bacteriophage-mediated manipulation of the gut microbiome\u2014Promises and presents limitations","volume":"44","author":"Rasmussen","year":"2020","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"01187","DOI":"10.3389\/fmicb.2019.01187","article-title":"Bacteriophage production models: An overview","volume":"10","author":"Latz","year":"2019","journal-title":"Front. Microbiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3869","DOI":"10.1128\/IAI.73.7.3869-3877.2005","article-title":"Listeriolysin O-induced membrane permeation mediates persistent interleukin-6 production in Caco-2 cells during Listeria monocytogenes infection in vitro","volume":"73","author":"Tsuchiya","year":"2005","journal-title":"Infect. Immun."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Kutter, E., and Sulakvelidze, A. (2004). Bacteriophage therapy in humans. Bacteriophages: Biology and Applications, CRC Press.","DOI":"10.1201\/9780203491751.ch14"},{"key":"ref_42","first-page":"211","article-title":"Measurement of gastrointestinal pH and regional transit times in normal children","volume":"11","author":"Fallingborg","year":"1990","journal-title":"J. Pediatr. Gastroenterol. Nutr."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"446","DOI":"10.1128\/AAC.00635-06","article-title":"Efficacy of bacteriophage therapy against gut-derived sepsis caused by Pseudomonas aeruginosa in mice","volume":"51","author":"Watanabe","year":"2007","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_44","first-page":"51","article-title":"The factors affecting effectiveness of treatment in phages therapy","volume":"5","year":"2014","journal-title":"Front. Microbiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1016\/j.chom.2020.06.002","article-title":"The Spatial Heterogeneity of the Gut Limits Predation and Fosters Coexistence of Bacteria and Bacteriophages","volume":"28","author":"Chaffringeon","year":"2020","journal-title":"Cell Host Microbe"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.cis.2017.05.014","article-title":"Formulation, stabilisation and encapsulation of bacteriophage for phage therapy","volume":"249","author":"Malik","year":"2017","journal-title":"Adv. Colloid Interface Sci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"01984","DOI":"10.3389\/fmicb.2019.01984","article-title":"Bacteriophages Reduce Pathogenic Escherichia coli Counts in Mice Without Distorting Gut Microbiota","volume":"10","author":"Dissanayake","year":"2019","journal-title":"Front. Microbiol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/2\/190\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:04:07Z","timestamp":1760133847000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/2\/190"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,19]]},"references-count":47,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["v14020190"],"URL":"https:\/\/doi.org\/10.3390\/v14020190","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1,19]]}}}