Występowanie bakterii fermentacji mlekowej aktywnych względem Listeria w serach regionalnych wytwarzanych w okręgu tatrzańsko-beskidzkim

• Autorzy: Sip A. , Wieckowicz M. , Olejnik-Schmidt A. , Gardo A. , Gorlas R. , Grajek W.

Warianty tytułu

EN

Occurrence of lactic acid bacteria with activity against Listeria in Polish regional cheeses produced in the Tatrzansko-Beskidzki district

• Języki publikacji: PL

Abstrakty

PL

Celem pracy było wyizolowanie z serów regionalnych bakterii fermentacji mlekowej oraz przeprowadzenie ich selekcji pod kątem zdolności do antagonistycznego oddziaływania na Listeria. Dokonano również wstępnej charakterystyki czynników odpo­wiedzialnych za antagonizm względem Listeria oraz zidentyfikowano bakterie o najsil­niejszej listeriobójczej aktywności. Bakterie te przebadano ponadto w kierunku obecności genów bakteriocyn klasy Ila (bakteriocyn listeriobójczych). Materiałem izolacyjnym było pięć rodzajów serów regionalnych: bundz, bryndza, gołka niewędzona, gołka wędzona oraz warkocze. Jako podłoża izolacyjne stosowano pożywkę MRS i M17. Antylisteryj- ną aktywność pozyskanych izolatów oznaczano metodą punktowo-dyfuzyjną względem bakterii Listeria innocua. Identyfikacji wybranych bakterii fermentacji mlekowej doko­nywano metodą analizy sekwencji genu kodującego 16S rRNA, a geny bakteriocyn kla­sy Ila wykrywano, stosując zdegenerowane startery oligonukleotydowe. W wyniku prac izolacyjnych pozyskano 1000 czystych kultur bakterii fermentacji mlekowej. Co trzecia z nich wykazywała antagonistyczną aktywność względem Listeria. Najwięcej bakterii zdolnych do antylisteryjnego działania wykryto w obrębie populacji LAB wyizolowanych z bryndzy i gołki niewędzonej. Wśród kultur aktywnych wobec Listeria dominowały kultury o działaniu bakteriobójczym. Działanie to było przede wszystkim następstwem konkurencji oraz syntezy kwasów organicznych i H2O2. Jedynie niecały 1% badanych kultur, oprócz wymienionych związków, syntezował także inne metabolity o działaniu antylisteryjnym. Były nimi bakteriocyny. Zdolność do ich syntezy posiadały bakterie, które zidentyfikowano jako Lactococcus garvieae, Leuconostoc mesenteroides oraz Lactobacillus plantarum.

EN

The main objective of this study was the isolation and selection of lactic acid bacteria capable of showing antagonistic activity against Listeria. The main factors contrib­uting to such activity were examined and characterized and strains with superior antiliste- rial activity were selected and identified as well. Furthermore, a PCR analysis of genomic DNA isolated from strains with relevant antilisterial activity was performed in order to determine, whether these bacteria are capable of producing class IIa antilisterial bacterio- cins. Five different polish artisanal cheeses were chosen as a source of isolation of baterial strains: bundz, bryndza, non-smoked golka, smoked golka and warkocze. The isolated mi­croorganisms were primarily cultivated on MRS and M17 media. The antilisterial activity was evaluated by diffusion tests against an indicator strain - Listeria innocua. Phylogenetic identification of the strains with highest antilisterial activity was performed by 16S rDNA sequencing. The presence of class IIa bacteriocin coding gene within the genome of se­lected strains was detected using a panel of degenerate PCR primers. About 1000 cultures of lactic acid bacteria were obtained during the studies presented in this paper. Nearly one third of these cultures showed an antagonistic activity against Listeria. The greatest number of active strains were among the lactic acid bacteria isolated from bryndza and non-smoked golka and these strains were able to synthetize mainly hydrogen peroxide and organic acids. Only less than 1% of examined bacteria showed the ability of synthesizing other metabo­lites with antilisterial activity - the bacteriocins. Strains identified as Lactococcus garvieae, Leuconostoc mesenteroides and Lactobacillus plantarum showed the ability to synthetize such metabolites. Furthermore, the presence of class Ila bacteriocin coding gene within the genomic DNA of these bacteria was also detected. The structure, modes of action and biosynthesis pathways of bacteriocins synthetized by Lc. garvieae, Leu. mesenteroides and Lb. plantarum are still examined by the authors.

Słowa kluczowe

PL

Polska; zywnosc regionalna; sery; Tatry; Beskidy; Listeria monocytogenes; bakterie kwasu mlekowego; aktywnosc antybakteryjna; bakteriocyny

EN

Poland; regional food; cheese; Tatras Mountains; Beskids Mountains; Listeria monocytogenes; lactic acid bacteria; antibacterial activity; bacteriocin

• Wydawca: -
• Czasopismo: Acta Scientiarum Polonorum. Biotechnologia
• Rocznik: 2009
• Tom: 08
• Numer: 2
• Opis fizyczny: s.27-44,rys.,tab.,fot.,bibliogr.

Twórcy

autor

Sip A.

• Uniwersytet Przyrodniczy w Poznaniu, ul.Wojska Polskiego 48, 60-627 Poznań

autor

Wieckowicz M.

autor

Olejnik-Schmidt A.

autor

Gardo A.

autor

Gorlas R.

autor

Grajek W.

Bibliografia

• Abee T., Krockel L., Hill C., 1995. Bacteriocins, modes of action and potentials in food preserva­tion and control of food poisoning. Int. J. Food Microbiol., 28, 169-185.
• Alvarado C., Garcia-Almendarez B.E., Martin S.E., Regalado C., 2005. Anti-Listeria monocyto­genes bacteriocin-like inhibitory substances from Enterococcus faecium UQ31 isolated from artisan mexican-style cheese. Current Microbiol., 51, 110-115.
• Belgacem Z.B., Ferchichi M., Prevost H., Dusset X., Monai M., 2008. Screening for anti-listerial bacteriocin-producing lactic acid bacteria from "gueddid" a traditionally Tunisian fermented meat. Meat Sci., 78, 513-521.
• Cagiran H., 2004. Biotyping of Lactococcus garvieae isolated from Tyrkey. J. Fish. Aquatic Sci., 21, 267-269.
• Casia D., Requena T., Gomez R., 1996. Antimicrobial activity of lactic acid bacteria isolated from goat's milk and artisanal cheeses: characterization of a bacteriocin produced by Lactococcus curvatus IFPL105. J. Appl. Microbiol., 81, 35-41.
• CDC, 2000. Multistate outbreak of listeriosis - United States, 2000. MMWR Morb. Mortal. Wkly. Rep., 49, 1129-1130.
• Cleveland J., Montville T.J., Nes I.F., Chikindas M.L., 2001. Bacteriocins, safe, natural antimicro­bials for food preservation. Int. J. Food Microbiol., 71, 1-20.
• Cole M., Jones M., Holyoak C., 1990. The effect of pH, salt concentration and temperature on the survival and growth of Listeria monocytogenes. J Appl. Bacteriol., 69, 63-69.
• Cordano A.M., Rocourt J., 2001. Occurrence of Listeria monocytogenes in food in Chile. Int. J. Food Microbiol., 70, 175-178.
• Daeschel M.A., 1989. Antimicrobial substances from lactic acid bacteria for use as food preservatives. Food Technol., 1, 164-167.
• De Vuyst L., 1994. Bacteriocins produced by Lactococcus lactis strains, [w:] Bacteriocins of lactic acid bacteria. Ed. De Vuyst L I Vandamme E, London 1994, 143-149.
• Doganay M., 2003. Listeriosis, clinical presentation. FEMS Immunol. Medic. Microbiol., 35, 173­175.
• Durmaz H., 2008. Fate of Listeria monocytogenes in Carra cheese during manufacture and ripe­ning. J. Food Safety, 29, 253-260.
• Eldat A., Ghittino C., Asanta L., Bozzetta E., Goria M., Prearo M., Bercovier H., 1996. Enterococ- cus serolicida is a junior synonym of Lactococcus garvieae, a causative agent of septemia and meningoencephalitis in fish. Current Microbiol., 32, 85-88.
• Elliot J.A., Collins M.D., Pigott N.E., Facklam R.R., 1991. Differentiation of Lactococcus lactis and Lactococcus garvieae from humans by comparison of whole-cell protein patterns. J. Cli. Microbiol., 29, 2731-2734.
• Ennahar S., Assobhei O., Hasselmann C., 1996. Inhibition of Listeria monocytogenes in a smearsurface soft cheese by Lactobacillusplantarum WHE92. J. Food Protect., 61, 186-191.
• Ennahar S., Sashihara T., Sonomoto K., Ishizaki A., 2000. Class IIa bacteriocins: biosynthesis, structure and activity. FEMS Microbiol Rev., 24, 85-106.
• Ennahar S., Sonomoto K, Ishizaki A., 1999. Class IIa bacteriocins from lactic acid bacteria, anti­bacterial activity and food preservation. J. Biosci. Bioeng., 87, 705-716.
• Faber J.M., Peterkin P.I., 1991. Listeria monocytogenes, a foodborne pathogen. Microbiol Rev., 55, 476-511.
• Ferchichi M., Fere J., Mabrouk K., Monai M., 2001: Lactococcin MMFII, a novel class IIa bacteriocin produced by Lactococcus lactis MMFII, isolated from a Tunisian dairy product. FEMS Microbiol. Letters, 205, 49-55.
• Galvez A., Abriouel H., Lopez R.L., Omar N.B., 2007. Bacteriocin-based strategies for food bio- preservation. Int. J. Food Microbiol., 120, 51-70.
• Gameiro N., Ferreira-Dias S., Ferreira M., Brito L., 2007. Evolution of Listeria monocytogenes populations during the ripening of naturally contaminated raw ewe's milk cheese. Food Con­trol, 18, 1258-1262.
• Gandhi M., Chikindas M. L., 2007. Listeria, a foodborne pathogen that knows how to survive. Int. J. Food Microbiol., 113, 1-15.
• Grajek W., Sip A., 2004. Biologiczne utrwalanie żywności z wykorzystaniem metabolitów bakterii mlekowych, [w:] Bakterie fermentacji mlekowej. Klasyfikacja, metabolizm, genetyka, wyko­rzystanie. Red. Libudzisz Z., Wyd. Politechniki Łódzkiej, 175-226.
• Guerra M.M., McLauchlin J.M., Bernardo F.A., 2001. Listeria in ready to eat unprocessed foods produced in Portugal. Food Microbiol., 18, 423-429.
• Gurira O.Z., Buys E.M., 2005. Characterization and antimicrobial activity of Pediococcus species isolated from South African farm-style cheese. Food Microbiol., 22, 159-168.
• Hansen J.N., 1994. Nisin as a model food preservative. Crit. Rev. Food Sci. Nutr., 34, 69-93.
• Kav K., Erganis O., 2008. Antibiotic susceptibility ofLactococcus garvieae in rainbow trout farms. Biuletyn Inst. Wet. Puławy, 52, 223-226.
• Little C.L., Rhoades J.R., Sago S.K., Harris J., Greenwood M., Mithani V., Grant K., McLauchlin J., 2008. Microbiological quality of retail cheese made from raw, thermized or pasteurized milk in UK. Food Microbiol., 25, 304-312.
• Liu D., 2008. Epidemiology, [in:] Handbook of Listeria monocytogenes. Red. Liu D. CRC Press, New York, 27-60.
• Loncarevic S., Danielsson-Tham M.L., Tham W., 1995. Occurrence of Listeria monocytogenes in soft and semisoft cheeses in retail outlets in Sweden. Int. J. Food Microbiol., 26, 245-250.
• Lovett J., 1989. Listeria monocytogenes, [in:] Foodborne bacterial pathogens. Marcel Dekker, New York, 283-310.
• McLauchlin J., Mitchell R.T., Smerdon W.J., Jewell J., 2004. Listeria monocytogenes and listerio- sis, a hazard characterization for use in microbiological risk assessment of food. Int. J. Food Microbiol., 92, 15-33.
• Mead P.S. Slutsker L., Dietz V., Mccaig L. F., Bresse J.S., Shapiro C., Griffin P.M., Tauxe R.V., 1999. Food-related illness and death in United States. Emerg. Infect. Dis., 5, 607-625.
• Morgan S., Rossand R.P., Hill C., 1995. Bacteriolytic activity caused by the presence of a nov­el lactococcal plasmid encoding lactococcin A, B and M. Appl. Enviromen. Microbiol., 61, 2995-3001.
• Noves E., Silva A.C., Roche S.M., Velge P., Brito L., 2008. Virulence of Listeria monocytogenes isolated from the cheese dairy environment, other foods and clinical cases. J. Med. Microbiol., 57, 411-415.
• O'Sullivan L., Ross R.P., Hill C., 2002. Potential of bacteriocin-producing lactic acid bacteria for improvements in food safety and quality. Biochemie, 84, 593-604.
• Okutani A., Yumiko O., Yamamoto S., Igimi S., 2004. Overview of Listeria monocytogenes con­tamination. Int. J. Food Microbiol., 93, 131-140.
• Orndorff P.E., Hamrick T.S., Smoak I.W., Havell E.A., 2006. Host and bacterial factors in listeriosis pathogenesis. Vet. Microbiol., 114, 1-15.
• Parente E., Brienza C., Moles M., Ricciardi A., 1995. A comparison of methods for the measure­ment of bacteriocin activity. J. Microbiol. Methods, 22, 95-108.
• Reviriego C., Fernandez A., Horn N., Rodriguez E., Martin M.L., Fernandez L., Rodriguez J.M., 2004. Production of pediocin PA-1, and coproduction of nisin A and pediocin PA-1, by Lacto­coccus lactis strains of dairy origin. Int. Dairy J., 15, 45-49.
• Rodrigues U.M., Aguirre M., Facklam R.R., Collins M.D., 1991. Specific and intraspecific molecular typing of lactococci based on polymorphism of DNA encoding rRNA. J. Appl. Bacteriol., 71, 509-516.
• Rodriguez E., Calzada J., Arques J.L., Rodriquez J.M., Nunez M., Medina M., 2005. Antimicrobial activity of pediocin-producing Lactococcus lactis on Listeria monocytogenes, Staphylococcus aureus and Escherichia coli O157:H7 in cheese. Int. Dairy J., 15, 51-57.
• Schlech W.F., 1996. Overview of listeriosis. Food Control, 4/5, 183-186.
• Schlech W.F., 2000. Foodborne listeriosis. Clin. Infect. Dis., 31, 770-775.
• Silva M.C.D, Almeida R.C.C., Alves M.A.O., Almeida P.F., 2003. Occurrence of Listeria spp. in critical points and the environment of Minas Frescal cheese processing. Int. J. Food Microbiol., 81, 241-248.
• Silva M.C.D., Hofer E., Tibana A., 1998. Incidence of Listeria monocytogenes in cheese produced in Rio de Janeiro, Brazil. J. Food Protect., 61, 354-356.
• Sip A., Krasowska M., Więckowicz M., Grajek W., 2009. Metody skriningu bakteriocynogennych bakterii fermentacji mlekowej. Żywność. Nauka. Technologia. Jakość, 1, 5-26.
• Sip A., Więckowicz M., Krasowska M., 2009. Charakterystyka listeriobójczych bakteriocyn bakterii fermentacji mlekowej klasy IIa. Biotechnologia (w druku).
• Sorbino-Lopez A., Martin-Belloso O., 2008. Use of nisin and other bacteriocins for preservation of dairy products. Int. Dairy J., 18, 329-343.
• Tauxe R.V., 2002. Emerging foodborne pathogens. Int. J. Food Microbiol., 78, 31-41.
• Todorov S.D., Dicks L.M.T., 2006. Screening for bacteriocin-producing lactic acid bacteria from boza, a traditional cereal beverage from Bulgaria, comparision of the bacteriocins. Process Biochem., 41, 11-19.
• Todorov S.D., Nyati H., Meincken M., Dicks L.M.T., 2007. Partial characterization of bacteriocin AMA-K, produced by Lactobacillus plantarum AMA-K isolated from naturally fermented milk from Zimbabwe. Food Control, 18, 656-664.
• Topisirovic L., Kojic M., Fire D., Golic N., Strahinic I., Lozo J., 2006. Potential of lactic acid bacteria isolated from specific natural niches in food production and preservation. Int. J. Food Microbiol., 112, 230-235.
• Wagner M., McLauchlin J., 2008. Biology, [in:] Handbook of Listeria monocytogenes. Red. Liu D. CRC Press, New York, 3-26.
• Villani F., Aponte M., Blaiotta G., Maurello G., Pepe O., Moschetti G., 2002. Detection and characterization of a bacteriocin, garviecin L1-5, produced by Lactococcus garvieae isolated from raw crow's milk. J. Appl. Microbiol., 90, 430-439.