200 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /0, No..,,**,*2 (,**3) 8 Salmonella Enteritidis Inhibitors of Adhesion Ability of Salmonella Enteritidis

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1 200 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /0, No..,,**,*2 (,**3) 8 Salmonella Enteritidis Inhibitors of Adhesion Ability of Salmonella Enteritidis Takahisa Miyamoto, Joh Kawaguchi, Satoshi Shimotsu, Johtaro Kawagishi and Ken-Ichi Honjoh Laboratory of Food Hygienic Chemistry, Division of Food Biotechnology, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University, 0 +* +, Hakozaki, Higashi-ku, Fukuoka, 2+, 2/2+ Inhibitors of Salmonella Enteritidis adhesion to microplates were screened among -3 additives including natural and food additives known to be highly safe. S. Enteritidis adhered strongly to microplates after incubation in *+. Bacto-soytone for,. h. Adhesion was inhibited in the presence of pigments (Annatto pigment, Gardenia yellow, and Monascus pigment), food coloring products (Annatto AN, San-brown AC, San-yellow No., AU, San-red YM, San-red RCFU, and San-beet L), food additives (protamine, Chili extract, sucrose fatty acid esters, and monoglycerine fatty acid esters), and a flavonoid (Kaempferol). Among them, Annatto AN, Gardenia yellow, Monascus pigment, and sugar fatty acid esters inhibited adhesion at concentrations below antibacterial activity. In the case of sugar fatty acid esters of C2 to C +2, the longer the fatty acid chain the stronger the inhibitory e# ects. Monascus pigment and sucrose monostearate were the strongest inhibitors of S. Enteritidis adhesion, even at **+. without antibacterial activity. (Received Sep. 2,,**2 ; Accepted Dec.,.,,**2) Keywords : Salmonella, adhesion, inhibitor, sugar fatty acid ester, Monascus pigment : +* +** + +** - +33, +3 +, *- +/ +3 /. http : // 2+, 2/2+ 0 +* + Corresponding authortmiyamot@agr.kyushu-u.ac.jp, Salmonella Enteritidis. 1

2 9 : 201, S. Enteritidis S. Enteritidis LB / ml -* +-* rpm + *** +* L LB / ml -*,+ +-* rpm +- *** rpm / min,. OD +*. +** L 2 O +/1 /* L O+/1 +/* L 30 -* -1,. 3+* LB Trypticase Peptone Becton, Dickinson and Company,* Bacto-Soytone Difco Lacoratories Dried Yeast Extract D-- +,+ +/ D-( )-Glucose, D-( )- Fructose, D-( )-Galactose, D-( )-Mannose, D-( )-Lactose, D-( )-Mannitol, Xylitol, Maltose Monohydrate EB-DISK,/ *., m D-( )-Ra$ nose Pentahydrate +,+,* S. Enteritidis ++ - S. Enteritidis S. Enteritidis *+. Bacto-Soytone Difco Lacoratories OD *. 1 +/* L 30. EB-DISK,/ /* L S. Enteritidis -*,. S. Enteritidis -3-3 S. Enteritidis JA G S. Enteritidis ELA KA No. / No. -L AC AN No., AU MR YM RCFU L + Salmonella Enteritidis IFO --+- Luria Broth LB, Becton, Dickinson and Company 00* 00*

3 202 /0.,** , Mireles, +/ + +/* L,*. +/ 33,** L +** L /3/ nm Fig. + E# ects of components of medium for incubation Microplate Reader Model./* BIO-RAD on adhesion of S. Enteritidis onto the surface of microplate well / S. Enteritidis was incubated in the presence of Bactomicroplate Soytone ( ), Trypticase Peptone ( ), and Yeast extract / ( ) at various concentrations for,. h at -* in S. Enteritidis wells. After washing the plate wells, bac- terial cells adhered onto the plate wells were stained with crystal violet and measured. S. Enteritidis -*,. +* Table + +** L Tryptic S. Enteritidis Soy Agar TSA, Becton, Dickinson and Company -1.2, + S. Enteritidis S. Enteritidis + S. Enteritidis Bacto- Soytone *+. S. Enteritidis, S. Enteritidis S. Enteritidis IFO--*+ LB S, AN, Enteritidis -* -1 -* AC No., AU YM data not shown Fig. + RCFU L S. S. Enteritidis Enteritidis Trypticase Pep- Fig., tone Bacto-Soytone AN, Yeast extract *+. S. Enteritidis **+. Bacto-Soytone *+. +*. AN *+. S. Enteritidis **

4 11 : 203 Table + E# ects of sugars in incubation medium on adhesion of S. Enteritidis onto the surface of microplate well Absorbance at /3/ nm (MeanSD) Sugars Conc. * *4+* *4,* +4** Fructose *4+/, *4*+0 *4*2, *4*+0 *4*00 *4*+3 *4*.- *4*+, Glucose *4+-. *4*,+ *4+-3 *4*,0 *4+00 *4*+* *4+.0 *4**2 Galactose *4+/, *4*+0 *4+*1 *4**- *4+,0 *4*,, *4*0. *4**+ Mannose *4,-* *4*-* *4*11 *4**3 *4*1* *4*+- *4*12 *4*+. Mannitol *4,-* *4*-* *4+-+ *4*-1 *4*3+ *4*,* *4+,/ *4*/3 Xylitol *4+3+ *4*,* *4+1* *4*+2 *4+/3 *4*+. *4+10 *4*++ Lactose *4+23 *4**+ *4,-3 *4*,. *4,+* *4**- *4*0. *4**0 Maltose *4+23 *4**+ *4,++ *4**0 *4,,* *4*+- *4*2+ *4*+* Ra$ nose *4+30 *4*++ *4+0/ *4*++ *4+*1 *4**- *4*01 *4**. Soluble Starch *4+3+ *4*,* *4+03 *4*,* *4+.3 *4*+0 ND S. Enteritidis was incubated in the presence of various sugars at various concentrations for,. h at -* in microplate. After washing the plate wells, bacterial cells adhered onto the plate wells were stained with crystal violet and measured. Values are MeanSD for / separate experiments. ND : not done - +/ +, Fig S. Enteritidis S. Typhimurium +0 S. Enteritidis +2 *.*+ S. Enteritidis Table + **/. -+,+ +1 S. Enteritidis **+. Fig., S. Enteritidis - S. Enteritidis +.,* AN S. Enteritidis - twitching

5 204 /0.,**3. 12 Fig., E# ects of some natural colors and additives on adhesion and viability of S. Enteritidis onto the surface of microplate well and on viability S. Enteritidis was incubated in *+. Bacto-Soytone containing various natural colors and additives at various concentrations for,. h at -* in microplate. After washing the plate wells, S. Enteritidis adhered onto the plate wells were stained with crystal violet and measured. Values were the meansd for / separate experiments. Viable counts of the bacteria in the wells were counted by the plating method and the results were shown as an average of two separate experiments. ND : not done. S. Enteritidis S. Enteritidis HLB S. Enteritidis Hydro- phile-lipophile Balance ; HLB +3 S. Enteritidis HLB,+. 1 HLB S. Enteritidis

6 13 : 205 Fig. - E# ects of monoglycerine fatty acid esters on adhesion and viability of S. Enteritidis onto the surface of microplate well and on viability S. Enteritidis was incubated in *+. Bacto-Soytone containing monoglycerine fatty acid esters with di# erent fatty acids at various concentrations for,. h at -* in microplate. After washing the plate wells, bacterial cells adhered onto the plate wells were stained with crystal violet and measured. Values were the meansd for / separate experiments. Viable counts of the bacteria in the wells were counted by the plating method and the results were shown as an average of two separate experiments. ND : not done. AN /0,. S. Enteritidis

206 /0.,**3. 14 Fig.. E# ects of sucrose fatty acid esters on adhesion and viability of S. Enteritidis onto the surface of microplate well and on viability S. Enteritidis was incubated in *+.

7 206 /0.,**3. 14 Fig.. E# ects of sucrose fatty acid esters on adhesion and viability of S. Enteritidis onto the surface of microplate well and on viability S. Enteritidis was incubated in *+. Bacto-Soytone containing various sucrose fatty acid esters with di# erent fatty acids at various concentrations for,. h at -* in microplate. After washing the plate wells, bacterial cells adhered onto the plate wells were stained with crystal violet and measured. Values were the meansd for / separate experiments. Viable counts of the bacteria in the wells were counted by the plating method and the results were shown as an average of two separate experiments. ND : not done.,,,- HLB b-, +0

8 15 : 207 / 1,. **+. S. Enteritidis S. Enteritidis + +1 / ,**0 ready-to-eat, +/-.. Description of the dose-response relationship In,/,0 Microbiological Risk Assessment Series +, Risk Assessments of Salmonella in Eggs and Broiler Chickens, eds World Health Organization, Food and Agriculture Organization of the United Nations, (FAO/WHO), pp. -- -/ (,**,). -,3 +*1 ++.,**+. Solano, C., Garcia, B., Valle, J., Berasain, C., Ghigo, J.M., Gamazo, C. and Lasa, I., Genetic analysis of Salmonella enteritidis biofilm formation : critical role of cellulose. Mol. Microbiol.,.-, 13-2*2 (,**,). / Gerstel, U. and Romling, U., The csgd promoter, a con- trol unit for biofilm formation in Salmonella typhimurium. Research in Microbiology, +/., 0/3001 (,**-). 0 Collinson, S.K., Emody, L., Trust, T. J. and Kay, W.W., Thin aggregative fimbriae from diarrheagenic Escherichia S. Enteritidis coli. J. Bacteriol., +1.,..3*..3/ ( +33, ) Prigent-Combaret, C., Brombacher, E., Vidal, O., Ambert, A., Lejeune, P., Landini, P. and Dorel, A., Complex Regulatory Network Controls Initial Adhesion and Biofilm S. Enteritidis Formation in Escherichia coli via Regulation of the csgd Gene. J. Bacteriol., +2-, 1,+-1,,- (,**+). 2 AN AC NTS No., AU YM RCFU pp. +,- +-,,**2 L 3 Medina, M.B. and Fratamico, P.M., Binding interactions of collagen I, laminin and fibronectin with immobilized Escherichia coli O +/1 : H1 using a surface plasmon resonance biosensor. Biotechnology Techniques, +,,,-/,.* ( +332). AN +* Fratamico, P.M., Schultz, F. J., Benedict, R.C., Buchanan, R.L. and Cooke, P.H., Factors influencing attachment of Escherichia coli O +/1 : H1 to beef tissues and removal using selected sanitizing rinses. Journal of Food Protection, /3,./-./3 ( +330). ++ Miyamoto,T., Shimizu, Y., Kobayashi, H., Honjoh, K. and Iio, M., Studies of collagen binding with immobilized **+. S. Salmonella enteritidis and inhibition with synthetic and Enteritidis naturally occurring food additives by a surface plasmon resonance biosensor. Sensors and Materials, +/,./-.00 (,**-). +, Mireles, II, J., Toguchi, A. and Harshey, R.M., Salmonella +2 enterica serovar Typhimurium swarming mutants with altered biofilm-forming abilities : Surfactin inhibits biofilm formation. J. Bacteriol., +2-, /2./ /2/. (,**+). +- Jonas, K., Tomenius, H., Kader, A.,,, Normark, S., Römling, U., Belova, L.M. and Melefors,» O., Roles of curli,

9 208 /0.,**3. 16 cellulose and BapA in Salmonella biofilm morphology 22, +*.3 +*// ( +333). studied by atomic force microscopy, BMC Microbiology, +3, : 1* doi : +* / ,+2*-1-1* (,**1). This article is -3/,**+ available from : http : // ,*,+2*// 1 1*. / ,+,,2 0,-0--,*** -- -0/ -1,,**/ +/ Singh, P.K., Parsek, M.R., Greenberg, E.P. and Welsh, M.,, / + - J., A component of innate immunity prevents bacterial biofilm development Nature,.+1, //, /// (,**,). pp.,*0, Oyofo, B.A., Droleskey, R.E., Norman, J.O., Mollenhauer,,- / +. H.H., Ziprin, R.L., Corrier, D.E. and DeLoach, J.R. Inhibition by mannose of in vitro colonization of chicken pp.,++,-, +313 small intestine by Salmonella typhimurium. Poultry Sci.,,. /2+ 02, +-/+ +-/0 ( +323). +1 Potter, R., Hansen, T.L. and Gill, T.A. : Inhibition of pp...../ foodborne bacteria by native and modified protamine :,/ Importance of electrostatic interactions. Int. J. Food /-. +3,-,**- Microbiol., +*-,,--. (,**.).,0 +2 Hansen, L.T. and Gill, T.A. : Solubility and antimicrobial.0. J-,-1,-3,**/ e$ cacy of protamine on Listeria monocytogenes and Escherichia coli as influenced by ph. J. Appl. Microbiol.,,* 3 2,* +,,.

20 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /0, No. +,,* -* (,**3) 20 * * Taste of Mentsuyu (a Japanese noodle soup) Depends on the Combination and P

20 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /0, No. +,,* -* (,**3) 20 * * Taste of Mentsuyu (a Japanese noodle soup) Depends on the Combination and Proportion of Ingredients Akiko Otomi Keywords : taste,