Health Professions (miscellaneous); Microbiology; Food Science
Abstract :
[en] A Staphyloccoccus aureus is one of the leading causes of food poisoning outbreaks (FPOs) worldwide. Staphylococcal food poisoning (SFP) is induced by the ingestion of food containing sufficient levels of staphylococcal enterotoxins (SEs). Currently, 33 SEs and SE-like toxins (SEls) have been described in the literature, but only five named “classical” enterotoxins are commonly investigated in FPOs due to lack of specific routine analytical techniques. The aims of this study were to (i) establish the genetic profile of strains in a variety of artisanal cheeses (n = 30) in Belgium, (ii) analyze the expression of the SE(l)s by these strains and (iii) compare the output derived from the different analytical tools. Forty-nine isolates of S. aureus were isolated from ten Belgian artisanal cheeses and were analyzed via microbiological, immunological, liquid chromatography mass spectrometry, molecular typing and genetic methods. The results indicated that classical SEs were not the dominant SEs in the Belgian artisanal cheeses that were analyzed in this study, and that all S. aureus isolates harbored at least one gene encoding a new SE(l). Among the new SE(l)s genes found, some of them code for enterotoxins with demonstrated emetic activity and ecg-enterotoxins. It is worth noting that the involvement of some of these new SEs has been demonstrated in SFP outbreaks. Thus, this study highlighted the importance of the development of specific techniques for the proper investigation of SFP outbreaks.
Research center :
FARAH. Santé publique vétérinaire - ULiège
Disciplines :
Food science
Author, co-author :
Minutillo, Raphaëlle ✱; Université de Liège - ULiège > Fundamental and Applied Research for Animals and Health (FARAH) ; Department of Food Sciences, Fundamental and Applied Research for Animals & Health (FARAH), Faculty of Veterinary Medecine, Avenue de Cureghem 10, 4000 Liege, Belgium
Pirard, Barbara ✱; Université de Liège - ULiège > Fundamental and Applied Research for Animals and Health (FARAH) > FARAH: Santé publique vétérinaire ; Department of Food Sciences, Fundamental and Applied Research for Animals & Health (FARAH), Faculty of Veterinary Medecine, Avenue de Cureghem 10, 4000 Liege, Belgium
Fatihi, Abdelhak ; Laboratory for Food Safety, French Agency for Food, Environmental and Occupational Health & Safety (ANSES), Université Paris-Est, 94700 Maisons-Alfort, France
Cavaiuolo, Marina; Laboratory for Food Safety, French Agency for Food, Environmental and Occupational Health & Safety (ANSES), Université Paris-Est, 94700 Maisons-Alfort, France
Lefebvre, Donatien; Laboratory for Food Safety, French Agency for Food, Environmental and Occupational Health & Safety (ANSES), Université Paris-Est, 94700 Maisons-Alfort, France
Gerard, Amaury ; Université de Liège - ULiège > Département GxABT > Chemistry for Sustainable Food and Environmental Systems (CSFES) ; Brewing and Food Science Unit, LABIRIS, Avenue Emile Gryzon 1, 1070 Anderlecht, Belgium
Taminiau, Bernard ; Université de Liège - ULiège > Département de sciences des denrées alimentaires (DDA) > Microbiologie des denrées alimentaires ; Department of Food Sciences, Fundamental and Applied Research for Animals & Health (FARAH), Faculty of Veterinary Medecine, Avenue de Cureghem 10, 4000 Liege, Belgium
Nia, Yacine ; Laboratory for Food Safety, French Agency for Food, Environmental and Occupational Health & Safety (ANSES), Université Paris-Est, 94700 Maisons-Alfort, France
Hennekinne, Jacques-Antoine; Laboratory for Food Safety, French Agency for Food, Environmental and Occupational Health & Safety (ANSES), Université Paris-Est, 94700 Maisons-Alfort, France
Daube, Georges ; Université de Liège - ULiège > Département de sciences des denrées alimentaires (DDA) > Microbiologie des denrées alimentaires ; Department of Food Sciences, Fundamental and Applied Research for Animals & Health (FARAH), Faculty of Veterinary Medecine, Avenue de Cureghem 10, 4000 Liege, Belgium
Clinquart, Antoine ; Université de Liège - ULiège > Département de sciences des denrées alimentaires (DDA) > Technologie des denrées alimentaires ; Department of Food Sciences, Fundamental and Applied Research for Animals & Health (FARAH), Faculty of Veterinary Medecine, Avenue de Cureghem 10, 4000 Liege, Belgium
✱ These authors have contributed equally to this work.
Language :
English
Title :
The Enterotoxin Gene Profiles and Enterotoxin Production of Staphylococcus aureus Strains Isolated from Artisanal Cheeses in Belgium
Cardozo M.V. Nespolo N. Delfino T.C. Almeida C.C.D. Pizauro L.J.L. Valmorbida M.K. Pereira N. Ávila F.A.D. Raw milk cheese as a potential infection source of pathogenic and toxigenic food born pathogens Food Sci. Technol. 2021 41 355 358 10.1590/fst.37919
Viçosa G.N. Le Loir A. Le Loir Y. de Carvalho A.F. Nero L.A. Egc Characterization of enterotoxigenic Staphylococcus aureus isolates obtained from raw milk and cheese Int. J. Food Microbiol. 2013 165 227 230 10.1016/j.ijfoodmicro.2013.05.023 23800734
Asao T. Kumeda Y. Kawai T. Shibata T. Oda H. Haruki K. Nakazawa H. Kozaki S. An extensive outbreak of staphylococcal food poisoning due to low-fat milk in Japan: Estimation of enterotoxin A in the incriminated milk and powdered Skim Milk Epidemiol. Infect. 2003 130 33 40 10.1017/S0950268802007951
Le H.H.T. Dalsgaard A. Andersen P.S. Nguyen H.M. Ta Y.T. Nguyen T.T. Large-scale Staphylococcus aureus foodborne disease poisoning outbreak among primary school children Microbiol. Res. 2021 12 43 52 10.3390/microbiolres12010005
Kadariya J. Smith T.C. Thapaliya D. Staphylococcus aureus and staphylococcal food-borne disease: An ongoing challenge in public health BioMed Res. Int. 2014 2014 827965 10.1155/2014/827965
Abril G.A. Villa G.T. Barros-Velázquez J. Cañas B. Sánchez-Pérez A. Calo-Mata P. Carrera M. Staphylococcus aureus exotoxins and their detection in the dairy industry and mastitis Toxins 2020 12 537 10.3390/toxins12090537
Ertas N. Gonulalan Z. Yildirim Y. Kum E. Detection of Staphylococcus aureus enterotoxins in sheep cheese and dairy desserts by multiplex PCR technique Int. J. Food Microbiol. 2010 142 74 77 10.1016/j.ijfoodmicro.2010.06.002
Jakobsen R.A. Heggebø R. Sunde E.B. Skjervheim M. Staphylococcus aureus and Listeria monocytogenes in Norwegian raw milk cheese production Food Microbiol. 2011 28 492 496 10.1016/j.fm.2010.10.017
Spanu V. Scarano C. Cossu F. Pala C. Spanu C. De Santis E.P.L. Antibiotic resistance traits and molecular subtyping of Staphylococcus aureus isolated from raw sheep milk cheese: Staphylococcus aureus drug resistance J. Food Sci. 2014 79 M2066 M2071 10.1111/1750-3841.12590
Viçosa G.N. Moraes P.M. Yamazi A.K. Nero L.A. Enumeration of coagulase and thermonuclease-positive Staphylococcus spp. in raw milk and fresh soft cheese: An evaluation of Baird-Parker agar, rabbit plasma fibrinogen agar and the Petrifilm Staph Express Count System Food Microbiol. 2010 27 447 452 10.1016/j.fm.2009.12.007
Carneiro Aguiar R.A. Ferreira F.A. Dias R.S. Nero L.A. Miotto M. Verruck S. De Marco I. De Dea Lindner J. Graduate Student Literature Review: Enterotoxigenic potential and antimicrobial resistance of Staphylococci from Brazilian artisanal raw milk cheeses J. Dairy Sci. 2022 105 5685 5699 10.3168/jds.2021-21634
Lawrynowicz-Paciorek M. Kochman M. Piekarska K. Grochowska A. Windyga B. The Distribution of enterotoxin and enterotoxin-like genes in Staphylococcus aureus strains isolated from nasal carriers and food samples Int. J. Food Microbiol. 2007 117 319 323 10.1016/j.ijfoodmicro.2007.03.009 17467095
Gérard A. El-Hajjaji S. Burteau S. Fall P.A. Pirard B. Taminiau B. Daube G. Sindic M. Study of the microbial diversity of a panel of Belgian artisanal cheeses associated with challenge studies for Listeria monocytogenes Food Microbiol. 2021 100 103861 10.1016/j.fm.2021.103861
Blaiotta G. Marrone R. Aponte M. Peruzy M.F. Smaldone G. Vollano L. Murru N. Characterisation of conciato romano: One of the oldest Italian cheeses Int. Dairy J. 2021 120 105077 10.1016/j.idairyj.2021.105077
Rajkovic A. Staphylococcus: Food poisoning The Encyclopedia of Food and Health Academic Press Oxford, UK 2016 Volume 5 133 139 978-0-12-384953-3
Filipello V. Bonometti E. Campagnani M. Bertoletti I. Romano A. Zuccon F. Campanella C. Losio M.N. Finazzi G. Investigation and follow-up of a staphylococcal food poisoning outbreak linked to the consumption of traditional hand-crafted alm cheese Pathogens 2020 9 1064 10.3390/pathogens9121064
Necidová L. Šťástková Z. Pospíšilová M. Janštová B. Strejček J. Dušková M. Karpíšková R. Influence of soft cheese technology on the growth and enterotoxin production of Staphylococcus aureus Czech J. Food Sci. 2009 27 127 133 10.17221/131/2008-CJFS
Pelisser M.R. Klein C.S. Ascoli K.R. Zotti T.R. Arisi A.C.M. Ocurrence of Staphylococcus aureus and multiplex PCR detection of classic enterotoxin genes in cheese and meat products Braz. J. Microbiol. 2009 40 145 148 10.1590/S1517-83822009000100025
Akineden O. Hassan A.A. Schneider E. Usleber E. Enterotoxigenic properties of Staphylococcus aureus isolated from goats’ milk cheese Int. J. Food Microbiol. 2008 124 211 216 10.1016/j.ijfoodmicro.2008.03.027 18455257
Wieneke A.A. Roberts D. Gilbert R.J. Staphylococcal food poisoning in the United Kingdom, 1969–1990 Epidemiol. Infect. 1993 110 519 531 10.1017/S0950268800050949 8519317
Johler S. Giannini P. Jermini M. Hummerjohann J. Baumgartner A. Stephan R. Further evidence for staphylococcal food poisoning outbreaks caused by egc-encoded enterotoxins Toxins 2015 7 997 1004 10.3390/toxins7030997 25802973
Rola J.G. Korpysa-Dzirba W. Osek J. Prevalence of Staphylococcus aureus and staphylococcal enterotoxins at different stages of production of raw milk Cheeses—Preliminary results Bull. Vet. Inst. Pulawy 2013 57 341 345 10.2478/bvip-2013-0059
Teague N.S. Grigg S.S. Peterson J.C. Gómez G.A. Talkington D.F. Outbreak of staphylococcal food poisoning from a military unit lunch party—United States, July 2012 Morb. Mortal. Wkly. Rep. 2013 62 1026 1028
Bens C.C.P.M. Voss A. Klaassen C.H.W. Presence of a novel DNA methylation enzyme in methicillin-resistant Staphylococcus aureus isolates associated with pig farming leads to uninterpretable results in standard pulsed-field gel electrophoresis analysis J. Clin. Microbiol. 2006 44 1875 1876 10.1128/JCM.44.5.1875-1876.2006
Rabello R.F. Moreira B.M. Lopes R.M.M. Teixeira L.M. Riley L.W. Castro A.C.D. Multilocus sequence typing of Staphylococcus aureus isolates recovered from cows with mastitis in brazilian dairy herds J. Med. Microbiol. 2007 56 1505 1511 10.1099/jmm.0.47357-0
Sobral D. Schwarz S. Bergonier D. Brisabois A. Feßler A.T. Gilbert F.B. Kadlec K. Lebeau B. Loisy-Hamon F. Treilles M. et al. High Throughput multiple locus variable number of tandem repeat analysis (MLVA) of Staphylococcus aureus from human, animal and food sources PLoS ONE 2012 7 e33967 10.1371/journal.pone.0033967
Roussel S. Felix B. Vingadassalon N. Grout J. Hennekinne J.-A. Guillier L. Brisabois A. Auvray F. Staphylococcus aureus strains associated with food poisoning outbreaks in France: Comparison of different molecular typing methods, including MLVA Front. Microbiol. 2015 6 882 10.3389/fmicb.2015.00882
Merda D. Felten A. Vingadassalon N. Denayer S. Titouche Y. Decastelli L. Hickey B. Kourtis C. Daskalov H. Mistou M.-Y. et al. NAuRA: Genomic tool to identify staphylococcal enterotoxins in Staphylococcus aureus strains responsible for foodborne outbreaks Front. Microbiol. 2020 11 1483 10.3389/fmicb.2020.01483
Lefebvre D. Blanco-Valle K. Hennekinne J.-A. Simon S. Fenaille F. Becher F. Nia Y. Multiplex detection of 24 staphylococcal enterotoxins in culture supernatant using liquid chromatography coupled to high-resolution mass spectrometry Toxins 2022 14 249 10.3390/toxins14040249 35448858
Madahi H. Rostami F. Rahimi E. Safarpoor Dehkordi F. Prevalence of enterotoxigenic Staphylococcus aureus isolated from chicken nugget in Iran Jundishapur J. Microbiol. 2014 7 e10237 10.5812/jjm.10237
Hait J. Tallent S. Melka D. Keys C. Bennett R. Prevalence of enterotoxins and toxin gene profiles of Staphylococcus aureus isolates recovered from a bakery involved in a second staphylococcal food poisoning occurrence J. Appl. Microbiol. 2014 117 866 875 10.1111/jam.12571 24917203
Johler S. Weder D. Bridy C. Huguenin M.-C. Robert L. Hummerjohann J. Stephan R. Outbreak of staphylococcal food poisoning among children and staff at a swiss boarding school due to soft cheese made from raw milk J. Dairy Sci. 2015 98 2944 2948 10.3168/jds.2014-9123 25726108
Féraudet Tarisse C. Goulard-Huet C. Nia Y. Devilliers K. Marcé D. Dambrune C. Lefebvre D. Hennekinne J.-A. Simon S. Highly sensitive and specific detection of staphylococcal enterotoxins SEA, SEG, SEH, and SEI by immunoassay Toxins 2021 13 130 10.3390/toxins13020130 33572449
Umeda K. Ono H.K. Wada T. Motooka D. Nakamura S. Nakamura H. Hu D.-L. High production of egc2-related staphylococcal enterotoxins caused a food poisoning outbreak Int. J. Food Microbiol. 2021 357 109366 10.1016/j.ijfoodmicro.2021.109366
Dicks J. Turnbull J.D. Russell J. Parkhill J. Alexander S. Genome sequencing of a historic Staphylococcus aureus collection reveals new enterotoxin genes and sheds light on the evolution and genomic organization of this key virulence gene family J. Bacteriol. 2021 203 e00587-20 10.1128/JB.00587-20
Omoe K. Hu D.-L. Ono H.K. Shimizu S. Takahashi-Omoe H. Nakane A. Uchiyama T. Shinagawa K. Imanishi K. Emetic Potentials of newly identified staphylococcal enterotoxin-like toxins Infect. Immun. 2013 81 3627 3631 10.1128/IAI.00550-13
Ono H.K. Hirose S. Naito I. Sato’o Y. Asano K. Hu D.-L. Omoe K. Nakane A. The emetic activity of staphylococcal enterotoxins, SEK, SEL, SEM, SEN and SEO in a small emetic animal model, the house musk shrew Microbiol. Immunol. 2017 61 12 16 10.1111/1348-0421.12460
Zeaki N. Budi Susilo Y. Pregiel A. Rådström P. Schelin J. Prophage-encoded staphylococcal enterotoxin A: Regulation of production in Staphylococcus aureus strains representing different sea regions Toxins 2015 7 5359 5376 10.3390/toxins7124889
Varshney A.K. Mediavilla J.R. Robiou N. Guh A. Wang X. Gialanella P. Levi M.H. Kreiswirth B.N. Fries B.C. Diverse enterotoxin gene profiles among clonal complexes of Staphylococcus aureus isolates from the Bronx, New York Appl. Environ. Microbiol. 2009 75 6839 6849 10.1128/AEM.00272-09
Kläui A.J. Boss R. Graber H.U. Characterization and comparative analysis of the Staphylococcus aureus genomic island vSaβ: An in silico approach J. Bacteriol. 2019 201 e00777-18 10.1128/JB.00777-18
Schwendimann L. Berger T. Graber H.-U. Meier S. Hummerjohann J. Jakob E. Effect of scalding temperature on growth of Staphylococcus aureus and formation of staphylococcal enterotoxin during the production of alpine cheese in a laboratory cheesemaking Model J. Food Prot. 2020 83 1822 1828 10.4315/JFP-19-600
Bianchi D.M. Gallina S. Bellio A. Chiesa F. Civera T. Decastelli L. Enterotoxin gene profiles of Staphylococcus aureus isolated from milk and dairy products in Italy Lett. Appl. Microbiol. 2014 58 190 196 10.1111/lam.12182
Hwang S.Y. Kim S.H. Jang E.J. Kwon N.H. Park Y.K. Koo H.C. Jung W.K. Kim J.M. Park Y.H. Novel multiplex PCR for the detection of the Staphylococcus aureus superantigen and its application to raw meat isolates in Korea Int. J. Food Microbiol. 2007 117 99 105 10.1016/j.ijfoodmicro.2007.02.013
Grispoldi L. Karama M. Armani A. Hadjicharalambous C. Cenci-Goga B.T. Staphylococcus aureus enterotoxin in food of animal origin and staphylococcal food poisoning risk assessment from farm to table Ital. J. Anim. Sci. 2021 20 677 690 10.1080/1828051X.2020.1871428
Roque A.C.A. Lowe C.R. Taipa M.A. Antibodies and genetically engineered related molecules: Production and purification Biotechnol. Prog. 2004 20 639 654 10.1021/bp030070k 15176864
NF EN ISO 6887-5:2020 Microbiology of the Food Chain—Preparation of Test Samples, Initial Suspension and Decimal Dilutions for Microbiological Examination—Part 5: Specific Rules for the Preparation of Milk and Milk Products. International Organization for Standardization Geneva, Switzerland 2020
NF EN ISO 6888-2:1999 Microbiology of the Food Chain—Horizontal Method for Counting Positive Coagulase Staphylococci (Staphylococcus aureus and Other Species)—Part 2: Technique Using Rabbit Plasma and Fibrinogen Agar Medium International Organization for Standardization Geneva, Switzerland 1999
Kadiroğlu P. Korel F. Ceylan C. Quantification of Staphylococcus aureus in white cheese by the improved DNA extraction strategy combined with TaqMan and LNA Probe-Based qPCR J. Microbiol. Methods 2014 105 92 97 10.1016/j.mimet.2014.06.022 25016130
NF EN ISO 8261:2001 Files Provides General Guidelines for the Preparation of Test Samples, Primary Dilutions and Decimal Dilutions, for the Microbiological Examination of Milk and Dairy Products, Including Infant Formula Foods International Organization for Standardization Geneva, Switzerland 2001
Agence Nationale de la Sécurité Sanitaire Alomentation, Environnement, Travail (ANSES), Version 2—List of Methods Used in the Field of ANSES National Reference Laboratory’s Missions—Positive coagulase Staphylococci (PCS) Available online: https://www.anses.fr/fr/system/files/Liste%20des%20m%C3%A9thodes-LNR%20SCP.pdf (accessed on 12 June 2022)
Bankevich A. Nurk S. Antipov D. Gurevich A.A. Dvorkin M. Kulikov A.S. Lesin V.M. Nikolenko S.I. Pham S. Prjibelski A.D. et al. SPAdes: A New Genome Assembly Algorithm and Its Applications to Single-Cell Sequencing J. Comput. Biol. J. Comput. Mol. Cell Biol. 2012 19 455 477 10.1089/cmb.2012.0021
Geneious Biologics, Version 10.2.3 2023 Available online: https://www.geneious.com (accessed on 5 June 2023)
Benkerroum N. Staphylococcal enterotoxins and enterotoxin-like toxins with special reference to dairy products: An overview Crit. Rev. Food Sci. Nutr. 2018 58 1943 1970 10.1080/10408398.2017.1289149 28362113
Omoe K. Imanishi K. Hu D.-L. Kato H. Fugane Y. Abe Y. Hamaoka S. Watanabe Y. Nakane A. Uchiyama T. et al. Characterization of novel staphylococcal enterotoxin-like toxin type P Infect. Immun. 2005 73 5540 5546 10.1128/IAI.73.9.5540-5546.2005 16113270
Hennekinne J.-A. Guillier F. Perelle S. De Buyser M.-L. Dragacci S. Krys S. Lombard B. Intralaboratory validation according to the EN ISO 16 140 Standard of the Vidas SET2 detection Kit for use in official controls of staphylococcal enterotoxins in milk products J. Appl. Microbiol. 2007 102 1261 1272 10.1111/j.1365-2672.2006.03183.x
Vernozy-Rozand C. Mazuy-Cruchaudet C. Bavai C. Richard Y. Comparison of three immunological methods for detecting staphylococcal enterotoxins from food Lett. Appl. Microbiol. 2004 39 490 494 10.1111/j.1472-765X.2004.01602.x 15548300
Ono H.K. Hirose S. Narita K. Sugiyama M. Asano K. Hu D.-L. Nakane A. Histamine Release from Intestinal Mast Cells Induced by staphylococcal enterotoxin A (SEA) evokes vomiting reflex in common marmoset PLoS Pathog. 2019 15 e1007803 10.1371/journal.ppat.1007803
Bennett R.W. Staphylococcal enterotoxin and its Rapid identification in foods by enzyme-linked immunosorbent assay–based Methodology J. Food Prot. 2005 68 1264 1270 10.4315/0362-028X-68.6.1264 15954720
Derzelle S. Dilasser F. Duquenne M. Deperrois V. Differential temporal expression of the staphylococcal enterotoxins genes during cell growth Food Microbiol. 2009 26 896 904 10.1016/j.fm.2009.06.007
Collery M.M. Smyth D.S. Tumilty J.J.G. Twohig J.M. Smyth C.J. Associations between enterotoxin gene cluster types egc1, egc2 and egc3, agr types, enterotoxin and enterotoxin-like gene profiles, and molecular typing characteristics of human nasal carriage and animal isolates of Staphylococcus aureus J. Med. Microbiol. 2009 58 13 25 10.1099/jmm.0.005215-0
Omoe K. Ishikawa M. Shimoda Y. Hu D.-L. Ueda S. Shinagawa K. Detection of seg, seh, and sei genes in Staphylococcus aureus isolates and determination of the enterotoxin productivities of S. aureus isolates harboring seg, seh, or sei genes J. Clin. Microbiol. 2002 40 857 862 10.1128/JCM.40.3.857-862.2002
Chieffi D. Fanelli F. Cho G.-S. Schubert J. Blaiotta G. Franz C.M.A.P. Bania J. Fusco V. Novel insights into the enterotoxigenic potential and genomic background of Staphylococcus aureus isolated from raw milk Food Microbiol. 2020 90 103482 10.1016/j.fm.2020.103482
Mekhloufi O.A. Chieffi D. Hammoudi A. Bensefia S.A. Fanelli F. Fusco V. Prevalence, enterotoxigenic potential and antimicrobial resistance of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA) isolated from Algerian ready to eat foods Toxins 2021 13 835 10.3390/toxins13120835 34941673
Hummerjohann J. Naskova J. Baumgartner A. Graber H.U. Enterotoxin-producing Staphylococcus aureus Genotype B as a major contaminant in Swiss raw milk cheese J. Dairy Sci. 2014 97 1305 1312 10.3168/jds.2013-7643 24440268
