{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T16:11:49Z","timestamp":1790611909193,"version":"4.1.0"},"reference-count":52,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2022,11,30]],"date-time":"2022-11-30T00:00:00Z","timestamp":1669766400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Foods"],"abstract":"<jats:p>Bacteria enter milk during poor hygiene practices and can form a biofilm on surfaces that come into contact with human milk. The presence of a biofilm increases the risk of infections among newborns as bacteria protected by biofilm are resistant to washing and disinfection processes. The formation of the biofilm depends on the microbial species, environmental conditions, and the specific materials colonized. The aim of this study is to analyze the effects of factors such as temperature, incubation time, and initial cell concentration on biofilm formation by pathogenic bacteria isolated from human milk on model hydrophobic polystyrene surfaces. Model studies confirm that pathogenic bacteria appearing in human milk as a result of cross-contamination tend to form a biofilm. The majority of isolates formed biofilm at both 25 and 37 \u00b0C after 12 h at 1 \u00d7 103 CFU\/mL inoculum count. Multivariate principal component analysis (PCA) showed that at lower temperatures, biofilm formation by bacterial isolates was the main determinant of biofilm formation, other factors were less important; however, at 37 \u00b0C, time was a factor in biofilm formation. The model research performed underlines the importance of maintaining the proper hygiene of rooms, surfaces, and devices for expressing, storing, and preparing mothers\u2019 milk and powdered infant formula (PIF) in facilities responsible for feeding newborns and premature babies.<\/jats:p>","DOI":"10.3390\/foods11233862","type":"journal-article","created":{"date-parts":[[2022,11,30]],"date-time":"2022-11-30T03:49:13Z","timestamp":1669780153000},"page":"3862","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Factors Influencing Biofilm Formation by Salmonella enterica sv. Typhimurium, E. cloacae, E. hormaechei, Pantoea spp., and Bacillus spp. Isolated from Human Milk Determined by PCA Analysis"],"prefix":"10.3390","volume":"11","author":[{"given":"Mateusz","family":"Gemba","sequence":"first","affiliation":[{"name":"Institute of Human Nutrition, Department of Food Hygiene and Quality Management, Warsaw University of Life Sciences\u2014SGGW, 02-786 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8027-4914","authenticated-orcid":false,"given":"El\u017cbieta","family":"Rosiak","sequence":"additional","affiliation":[{"name":"Institute of Human Nutrition, Department of Food Hygiene and Quality Management, Warsaw University of Life Sciences\u2014SGGW, 02-786 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zuzanna","family":"Nowak-\u017byczy\u0144ska","sequence":"additional","affiliation":[{"name":"Institue of Animal Sciences, Departament of Food Hygiene and Quality Management, Warsaw University of Life Sciences\u2014SGGW, 02-768 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paulina","family":"Ka\u0142\u0119cka","sequence":"additional","affiliation":[{"name":"Institute of Human Nutrition, Department of Food Hygiene and Quality Management, Warsaw University of Life Sciences\u2014SGGW, 02-786 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"El\u017cbieta","family":"\u0141odykowska","sequence":"additional","affiliation":[{"name":"St. Holy Family Specialist Hospital, Regional Bank of Human Milk, 02-544 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Danuta","family":"Ko\u0142o\u017cyn-Krajewska","sequence":"additional","affiliation":[{"name":"Institute of Human Nutrition, Department of Food Hygiene and Quality Management, Warsaw University of Life Sciences\u2014SGGW, 02-786 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1016\/j.siny.2016.05.003","article-title":"The human milk microbiome and factors influencing its composition and activity","volume":"21","author":"Salminen","year":"2016","journal-title":"Semin. Fetal Neonatal Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.phrs.2012.09.001","article-title":"The human milk microbiota: Origin and potential roles in health and disease","volume":"69","author":"Langa","year":"2013","journal-title":"Pharmacol. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"761","DOI":"10.15585\/mmwr.mm6628a5","article-title":"Notes from the field: Cronobacter sakazakii infection associated with feeding extrinsically contaminated expressed human milk to a premature infant\u2014Pennsylvania, 2016","volume":"66","author":"Bowen","year":"2017","journal-title":"MMWR Morb. Mortal. Wkly. Rep."},{"key":"ref_4","first-page":"12","article-title":"Notes from the field: Cronobacter sakazakii meningitis in a full-term neonate fed exclusively with breast milk\u2014Indiana, 2018","volume":"67","author":"Sundararajan","year":"2018","journal-title":"Morb. Mortal. Wkly. Rep."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"393","DOI":"10.3201\/eid2402.171411","article-title":"Cronobacter sakazakii infection from expressed breast milk, Australia","volume":"24","author":"McMullan","year":"2018","journal-title":"Emerg. Inf. Dis."},{"key":"ref_6","first-page":"1","article-title":"Klebsiella pneumoniae in breast milk-A cause of sepsis in neonate","volume":"9","author":"Pawlik","year":"2017","journal-title":"Arch. Med."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Cormontagne, D., Rigourd, V., Vidic, J., Rizzotto, F., Bille, E., and Ramarao, N. (2021). Bacillus cereus Induces Severe Infections in Preterm Neonates: Implication at the Hospital and Human Milk Bank Level. Toxins, 13.","DOI":"10.3390\/toxins13020123"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1016\/j.ajic.2013.01.043","article-title":"Bacillus cereus and severe intestinal infections in preterm neonates: Putative role of pooled breast milk","volume":"41","author":"Decousser","year":"2013","journal-title":"Am. J. Infect. Control"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1089\/bfm.2015.0146","article-title":"Mother\u2019s Milk as a Source of Enterobacter cloacae Sepsis in a Preterm","volume":"10","author":"Weems","year":"2015","journal-title":"Infant. Breastfeed. Med."},{"key":"ref_10","unstructured":"Food Agricultural Organization, and World Health Organization (2004). Enterobacter sakazakii and other microorganisms in powdered infant formula. Meeting Report Microbiological Risk Assessment 6, WHO."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Doijad, S.P., Barbuddhe, S.B., Garg, S., Poharkar, K.V., Kalorey, D.R., Kurkure, N.V., Rawool, D.B., and Chakraborty, T. (2015). Biofilm-forming abilities of Listeria monocytogenes serotypes isolated from different sources. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0137046"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1128\/IAI.05322-11","article-title":"Molecular Characterization of UpaB and UpaC, Two New Autotransporter Proteins of Uropathogenic Escherichia coli CFT073","volume":"80","author":"Allsopp","year":"2012","journal-title":"Infect. Immun."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.ijfoodmicro.2010.02.005","article-title":"Differences in attachment of Salmonella enterica serovars to cabbage and lettuce leaves","volume":"139","author":"Patel","year":"2010","journal-title":"Int. J. Food Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/j.ijfoodmicro.2009.08.007","article-title":"Biofilm formation on enteral feeding tubes by Cronobacter sakazakii, Salmonella serovars and other Enterobacteriaceae","volume":"136","author":"Hurrell","year":"2009","journal-title":"Int. J. Food Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/S0924-2244(03)00155-9","article-title":"Risk profile of Enterobacter sakazakii, an emergent pathogen associated with infant milk formula","volume":"14","author":"Iversen","year":"2003","journal-title":"Trends Food Sci. Technol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5783","DOI":"10.1080\/10408398.2021.1892585","article-title":"Review of multi-species biofilm formation from foodborne pathogens: Multi-species biofilms and removal methodology","volume":"62","author":"Kim","year":"2022","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Carrascosa, C., Raheem, D., Ramos, F., Saraiva, A., and Raposo, A. (2021). Microbial Biofilms in the Food Industry\u2014A Comprehensive Review. Int. J. Environ. Res. Public Health, 18.","DOI":"10.3390\/ijerph18042014"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"4014","DOI":"10.1002\/jsfa.8945","article-title":"Salmonella and Campylobacter biofilm formation: A comparative assessment from farm to fork","volume":"98","author":"Lamas","year":"2018","journal-title":"J. Sci. Food Agric."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"0036850419898659","DOI":"10.1177\/0036850419898659","article-title":"Environmental factors modulate biofilm formation by Staphylococcus aureus","volume":"103","author":"Liu","year":"2020","journal-title":"Sci. Prog."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1016\/j.fm.2009.09.009","article-title":"Cross-contamination of fresh-cut lettuce after a short-term exposure during pre-washing cannot be controlled after subsequent washing with chlorine dioxide or sodium hypochlorite","volume":"27","author":"Gil","year":"2010","journal-title":"Food Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1016\/j.fm.2014.04.015","article-title":"Biofilm-associated persistence of food-borne pathogens","volume":"45","author":"Bridier","year":"2015","journal-title":"Food Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e12430","DOI":"10.1111\/jfs.12430","article-title":"Biofilm formation and genetic characterization of New Zealand Cronobacter isolates","volume":"38","author":"Gupta","year":"2018","journal-title":"J. Food Saf."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"982","DOI":"10.1126\/science.1211037","article-title":"Active starvation responses mediate antibiotic tolerance in biofilms and nutrient-limited bacteria","volume":"334","author":"Nguyen","year":"2011","journal-title":"Science"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1016\/j.jhin.2011.08.013","article-title":"Effect of biofilm formation on the survival of Acinetobacter baumannii on dry surfaces","volume":"80","author":"Espinal","year":"2012","journal-title":"J. Hosp. Infect."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1016\/j.foodcont.2015.08.027","article-title":"Sporulation dynamics and spore heat resistance in wet and dry biofilms of Bacillus cereus","volume":"60","author":"Hayrapetyan","year":"2016","journal-title":"Food Control"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.1111\/jam.13533","article-title":"Biofilm formation and antibiotic resistance in Klebsiella pneumoniae urinary strains","volume":"123","author":"Vuotto","year":"2017","journal-title":"J. App. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.chom.2019.06.002","article-title":"Surviving as a Community: Antibiotic Tolerance and Persistence in Bacterial Biofilms","volume":"26","author":"Yan","year":"2019","journal-title":"Cell Host Microbe"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Zhu, T., Yang, C., Bao, X., Chen, F., and Guo, X. (2022). Strategies for controlling biofilm formation in food industry. Grain Oil Sci. Technol., 1\u201314.","DOI":"10.1016\/j.gaost.2022.06.003"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/2110-5820-1-19","article-title":"Hacking into bacterial biofilms: A new therapeutic challenge","volume":"1","author":"Bordi","year":"2011","journal-title":"Ann. Intensive Care"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1111\/j.1472-765X.2004.01591.x","article-title":"Attachment and biofilm formation on stainless steel by Escherichia coli O157: H7 as affected by curli production","volume":"39","author":"Ryu","year":"2004","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1007\/s12223-021-00864-2","article-title":"Bacterial biofilm formation on stainless steel in the food processing environment and its health implications","volume":"66","author":"Dula","year":"2021","journal-title":"Folia Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"111163","DOI":"10.1016\/j.foodres.2022.111163","article-title":"Antibiofilm efficacy of Leuconostoc mesent eroides J.27-derived postbiotic and food-grade essential oils against Vibrio parahaemolyticus, Pseudomonas aeruginosa, and Escherichia coli alone and in combination, and their application as a green preservative in the seafood industry","volume":"156","author":"Toushik","year":"2022","journal-title":"Food Res. Int."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1016\/S0740-0020(02)00123-5","article-title":"Influence of the incubation temperature, atmosphere and dynamic conditions on biofilm formation by Salmonella spp.","volume":"20","year":"2003","journal-title":"Food Microbiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1089\/fpd.2013.1710","article-title":"Ability of Salmonella spp. to produce biofilm is dependent on temperature and surface material","volume":"11","author":"Kaneno","year":"2014","journal-title":"Foodborne Pathog. Dis."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1875","DOI":"10.4315\/0362-028X.JFP-15-085","article-title":"Influence of temperature, source, and serotype on biofilm formation of Salmonella enterica isolates from pig slaughterhouses","volume":"78","author":"Piras","year":"2015","journal-title":"J. Food Protect."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1590\/1678-5150-pvb-4928","article-title":"Biofilm formation capacity of Salmonella serotypes at different temperature conditions","volume":"38","author":"Borges","year":"2018","journal-title":"Pesqui. Vet. Bras."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.ijfoodmicro.2018.05.011","article-title":"Predicting adhesion and biofilm formation boundaries on stainless steel surfaces by five Salmonella enterica strains belonging to different serovars as a function of pH, temperature and NaCl concentration","volume":"281","author":"Moraes","year":"2018","journal-title":"Int. J. Food Microbiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/j.fm.2017.03.005","article-title":"Microbiological quality of selected ready-to-eat leaf vegetables, sprouts and non-pasteurized fresh fruit-vegetable juices including the presence of Cronobacter spp.","volume":"65","author":"Garbowska","year":"2017","journal-title":"Food Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1870","DOI":"10.1093\/molbev\/msw054","article-title":"MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets","volume":"33","author":"Kumar","year":"2016","journal-title":"Mol. Biol. Evol."},{"key":"ref_40","first-page":"265","article-title":"Outbreak of a novel Enterobacter sp. Carrying blaCTX-M-15 in neonatal unit of a tertiary care hospital in Tanzania","volume":"38","author":"Mashana","year":"2011","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1297","DOI":"10.1007\/s10096-018-3249-z","article-title":"Recent actuality about Bacillus cereus and human milk bank: A new sensitive method for microbiological analysis of pasteurized milk","volume":"37","author":"Rigourd","year":"2018","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1186\/0717-6287-47-63","article-title":"Stress tolerant virulent strains of Cronobacter sakazakii from food","volume":"47","author":"Fakruddin","year":"2014","journal-title":"Biol. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"132","DOI":"10.4315\/0362-028X-51.2.132","article-title":"Thermal destruction of Escherichia coli and Klebsiella pneumoniae in human milk","volume":"51","author":"Morgan","year":"1988","journal-title":"J. Food Prot."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"012053","DOI":"10.1088\/1755-1315\/107\/1\/012053","article-title":"Biofilm formation as a method of survival of Escherichia coli and Pantoea spp in the marine environment","volume":"107","author":"Buzoleva","year":"2018","journal-title":"IOP Conf. Ser. Earth Environ. Sci."},{"key":"ref_45","first-page":"1","article-title":"Comparison of biofilm and attachment mechanisms of a phytopathological and clinical isolate of Klebsiella pneumoniae subsp. pneumoniae","volume":"10","author":"Korres","year":"2013","journal-title":"Sci. World J."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1007\/s12223-010-0012-y","article-title":"Temperature and pH affect the production of bacterial biofilm","volume":"55","year":"2010","journal-title":"Folia. Microbiol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"9582","DOI":"10.3390\/molecules18089582","article-title":"Evaluation of the effect of different growth media and temperature on the suitability of biofilm formation by Enterobacter cloacae strains isolated from food samples in South Africa","volume":"18","author":"Nyenje","year":"2013","journal-title":"Molecules"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1111\/j.1472-765X.2004.01507.x","article-title":"The growth profile, thermotolerance and biofilm formation of Enterobacter sakazakii grown in infant formula milk","volume":"38","author":"Iversen","year":"2004","journal-title":"Lett. App. Microbiol."},{"key":"ref_49","first-page":"814","article-title":"Cluster Differences in Antibiotic Resistance, Biofilm Formation, Mobility, and Virulence of Clinical Enterobacter cloacae Complex","volume":"13","author":"Liu","year":"2022","journal-title":"Front. Microbiol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1103","DOI":"10.1007\/s10068-017-0129-8","article-title":"Biofilm formation of Bacillus cereus under food-processing-related conditions","volume":"26","author":"Kwon","year":"2017","journal-title":"Food Sci. Biotechnol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.ijfoodmicro.2015.02.005","article-title":"Comparative analysis of biofilm formation by Bacillus cereus reference strains and undomesticated food isolates and the effect of free iron","volume":"200","author":"Hayrapetyan","year":"2015","journal-title":"Int. J. Food Microbiol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.lwt.2015.01.035","article-title":"Effects of culture conditions on the biofilm formation of Cronobacter sakazakii strains and distribution of genes involved in biofilm formation","volume":"62","author":"Ye","year":"2015","journal-title":"LWT-Food Sci. Technol."}],"container-title":["Foods"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2304-8158\/11\/23\/3862\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:29:57Z","timestamp":1760146197000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2304-8158\/11\/23\/3862"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,30]]},"references-count":52,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["foods11233862"],"URL":"https:\/\/doi.org\/10.3390\/foods11233862","relation":{},"ISSN":["2304-8158"],"issn-type":[{"value":"2304-8158","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,11,30]]}}}