Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR

TERZİ GÜLEL, GÖKNUR; gücükoğlu, ali; ÇADIRCI, Özgür; ALİŞARLI, MUSTAFA; KEVENK, Tahsin Onur

Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR

Özgür ÇADIRCI, (Ondokuz Mayıs Üniversitesi, Veteriner Fakültesi, Besin Hijyeni ve Teknolojisi Bölümü, Samsun,Türkiye)

Ali GÜCÜKOĞLU, (Ondokuz Mayıs Üniversitesi, Veteriner Fakültesi, Besin Hijyeni ve Teknolojisi Bölümü, Samsun,Türkiye)

GÖKNUR TERZİ GÜLEL, (Ondokuz Mayıs Üniversitesi, Veteriner Fakültesi, Besin Hijyeni ve Teknolojisi Bölümü, Samsun,Türkiye)

Tahsin Onur KEVENK, (Ondokuz Mayıs Üniversitesi, Veteriner Fakültesi, Besin Hijyeni ve Teknolojisi Bölümü, Samsun,Türkiye)

Mustafa ALİŞARLI (Ondokuz Mayıs Üniversitesi, Veteriner Fakültesi, Besin Hijyeni ve Teknolojisi Bölümü, Samsun,Türkiye)

Kafkas Üniversitesi Veteriner Fakültesi Dergisi

8 3

Yıl: 2013 Cilt: 19 Sayı: 5 Sayfa Aralığı: 869 - 874 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR

Öz:

Bu çalışmanın amacı sütlü tatlılarda Bacillus cereus varlığını araştırmak ve enterotoksijenik genleri multipleks PCR tekniğiyle belirlemektir. Bu amaçla 25 keşkül, 12 tavuk göğsü, 12 kazandibi, 30 supangle, 4 profiterol ve 17 sütlaç olmak üzere toplamda 100 örnek analiz edilmiştir. 3ü tavuk göğsü (%25), 2si supangle (%6.6), 1i keşkül (%4) ve 1i sütlaç (%5.8) olmak üzere toplamda 7 örnek (%7.0) B. cereus varlığı açısından pozitif bulunmuştur. Analiz edilen sütlü tatlı örneklerinde ortalama B. cereus sayısı 2.0×101 - 5.0×102 kob/g olarak tespit edilmiştir. 7 pozitif örnekten toplamda 20 izolat elde edilmiştir. Sonuçta, B. cereus izolatlarının %70inin (14/20) hbl geni taşımadığı, %30unun (6/20) üç enterotoksik HBL kompleksi kodlayıcı geni (hblA, hblC ve hblD) bulundurduğu saptanmıştır. Ayrıca yapılan incelemede, izolatların %40ının (8/20) üç NHE kompleksi kodlayıcı geni (nheA, nheB ve nheC), %45inin (9/20) iki nhe geni (nheA ve nheB) ve %15inin (3/20) bir adet nhe geni (ikisi nheA ve biri nheB) içerdiği saptanmıştır. ctyK1 geni hiçbir örnekte belirlenememiştir. Sütlü tatlılarda B. cereus varlığının gıda zehirlenmelerine yol açarak halk sağlığı açısından potansiyel bir tehlike olabileceği düşünülmektedir.

Anahtar Kelime:

Konular: Veterinerlik

Sütlü tatlılardan izole edilen Bacillus cereus suşlarının enterotoksijenik gen profillerinin multipleks PCR ile belirlenmesi

Öz:

The aim of this study was to investigate the presence of Bacillus cereus and to detect enterotoxigenic genes in dairy dessert samples utilising a multiplex PCR technique. A total of 100 samples, 25 keskul, 12 tavuk gogsu, 12 kazandibi, 30 supangle, 4 profiteroles and 17 sutlac were analysed. B. cereus contamination was found in 7 of 100 (7.0%) samples including 3 (25%) tavuk gogsu, 2 (6.6%) supangle, 1 (4%) keskul and 1 (5.8%) sutlac. The average number of B. cereus was between 2.0×101 - 5.0×102 cfu/g in these dairy dessert samples. A total of 20 isolates were collected from the 7 positive samples. The results indicated that 30% (6/20) of B. cereus isolates contain three enterotoxic HBL complex encoding genes hblA, hblC and hblD, whereas 70% (14/20) had no hbl genes. In addition, all three enterotoxic NHE complex encoding genes, nheA, nheB and nheC, were detected in 40% (8/20), two nhe genes (nheA and nheB) were found in 45% (9/20) and one nhe gene (two nheA and one nheB) was found in 15% (3/20) of the isolates. The ctyK1 gene was not detected in any sample. The presence of B. cereus and their enterotoxigenic genes in dairy desserts may be a potential risk for public health.

Anahtar Kelime:

Konular: Veterinerlik

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

1. Nakamura LK: Bacillus pseudomycoides sp. nov. Int J Syst Bacteriol, 48, 1031-1035, 1998.
2. Lechner S, Mayr R, Francis KP, Pruss BM, Kaplan T, Wiessner Gunkel E, Stewart GS, Scherer S: Bacillus weihenstephanensis sp. nov. is a new psychrotolerant species of the Bacillus cereus group. Int J Syst Bacteriol, 48, 1373-1382, 1998.
3. Kramer JM, Gilbert RJ: Bacillus cereus and other Bacillus species. In, Doyle MP (Ed): Foodborne Bacterial Pathogens. 21-70, Marcel Dekker Inc., New York, 1989.
4. Agata N, Mori M, Ohta M, Suwan S, Ohtani I, Isobe M: A novel dodecadepsipeptide, cereulide, isolated from Bacillus cereus causes vacuole formation in HEp-2 cells. FEMS Microbiol Lett, 121, 31-34, 1994.
5. Agata N, Ohta M, Arakawa Y, Mori M: The bceT gene of Bacillus cereus encodes an enterotoxic protein. Microbiology, 141, 983-988, 1995.
6. Beecher DJ, Wong ACL: Improved purification and characterization of hemolysin BL, a hemolytic dermonecrotic vascular permeability factor from Bacillus cereus. Infect Immunol, 62, 980-986, 1994.
7. Beecher DJ, Wong ACL: Tripartite hemolysin BL from Bacillus cereus. Hemolytic analysis of component interactions and a model for its characteristics paradoxical zone phenomenon. J Biol Chem, 272, 233-239, 1997.
8. Granum PE, OSullivan K, Lund T: The sequence of the non-haemolytic enterotoxin operon from Bacillus cereus. FEMS Microbiol Lett, 177, 225-229, 1999.
9. Christiansson A: Bacillus cereus. In, Roginsky H, Fuquay JW, Fox PF (Eds): Encyclopedia of Dairy Sciences. 123-128, Academic Press, Amsterdam, The Netherlands, 2003.
10. Beattie SH, Williams AG: Detection of toxigenic strains of Bacillus cereus and other Bacillus spp. with an improved cytotoxicity assay. Lett Appl Microbiol, 28, 221-225, 1999.
11. Granum PE, Brynestad S, Kramer JM: Analysis of enterotoxin production by Bacillus cereus from dairy products, food poisoning incidents and non-gastrointestinal infections. Int J Food Microbiol, 17, 269-279, 1993.
12. Sutherland AD: Toxin production by Bacillus cereus in dairy products. J Dairy Res, 60, 569-574, 1993.
13. Te Giffel MC, Beumer RR, Leijendekkers S, Rombouts FM: Incidence of Bacillus cereus and Bacillus subtilis in foods in the Netherlands. Food Microbiol, 13, 53-58, 1996.
14. Meer RR, Baker J, Bodyfelt FW, Griffiths MW: Psychrotrophic Bacillus spp. in fluid milk products. J Food Protect, 54, 969-979, 1991.
15. Christiansson A, Bertilsson J, Svensson B: Bacillus cereus spores in raw milk: factors affecting the contamination of milk during the grazing period. J Dairy Sci, 82, 305-314, 1999.
16. Torkar KG, Matijaic´ BB: Partial characterisation of bacteriocins produced by Bacillus cereus isolates from milk and milk products. Food Technol Biotech, 41, 121-129, 2003.
17. Iurlina MO, Saiz AM, Fuselli SR, Fritz R: Prevalence of Bacillus spp. in different food products collected in Argentina. LWT-Food Sci Technol, 39, 105-110, 2006.
18. Tham WA, Hadju LJ, Danielsson-Tham MLV: Bacteriological quality of on-farm manufactured goat cheese. Epidemiol Infect, 104, 87-100, 1990.
19. Pirttijarvi TS, Ahonen LM, Maunuksela LM, Salkinoja Salonen MS: Bacillus cereus in a whey process. Int J Food Microbiol, 44, 31-41, 1998.
20. Sarrias JA, Valero M, Salmeron MC: Enumeration, isolation and characterization of Bacillus cereus strains from Spanish raw rice. Food Microbiol, 19, 589-595, 2002.
21. Kursun O, Guner A, Ozmen G: Prevalence of Bacillus cereus in rabbit meat consumed in Burdur-Turkey, its enterotoxin producing ability and antibiotic susceptibility. Kafkas Univ Vet Fak Derg, 17 (Suppl A): S31-S35, 2011.
22. Wehrle E, Moravek M, Dietrich R, Bürk C, Didie A, Märtlbauer E: Comparison of multiplex PCR, enzyme immunoassay and cell culture methods for the detection of enterotoxinogenic Bacillus cereus. J Microbiol Meth, 78, 265-270, 2009.
23. Hanashiro A, Morita M, Matte GR, Matte MH, Torres EAFS: Microbiological quality of selected street foods from a restricted area of Sao Paulo city Brazil. Food Control, 16, 439-444, 2005.
24. Borge GIA, Skeie M, Sorhaug T, Langsrud T, Granum PE: Growth and toxin profiles of Bacillus cereus isolated from different food sources. Int J Food Microbiol, 69, 237-246, 2001.
25. ICMSF: International Commission on Microbiological Specification for Foods. Microorganisms in Foods, 1st ed., Clays Ltd., Suffolk, Great Britain, 1996.
26. Szabo RA, Todd ECD, Rayman MK: Twenty-four hours isolation and confirmation of Bacillus cereus in foods. J Food Protect, 47, 856-860, 1984.
27. Ahmed AA, Moustafa MK, Marth EH: Incidence of Bacillus cereus in milk and some milk products. J Food Protect, 46, 126-128, 1983.
28. Te Giffel MC, Beumer RR, Granum PE, Rombouts FM: Isolation and characterization of Bacillus cereus from pasteurised milk in household refrigerators in the Netherlands. Int J Food Microbiol, 34, 307-318, 1997.
29. Larsen HD, Jørgensen K: The occurrence of Bacillus cereus in Danish pasteurized milk. Int J Food Microbiol, 34, 179-186, 1997.
30. Ozdemir H: Pastörize sütlerde Bacillus cereusun varlığı. Gıda Dergisi, 28 (6): 611-614, 2003.
31. Molva C, Sudagidan S, Okuklu B: Extracellular enzyme production and enterotoxigenic gene profiles of Bacillus cereus and Bacillus thuringiensis strains isolated from cheese in Turkey. Food Control, 20, 829-834, 2009.
32. Gaviria A, Rivera M, Granum PE, Priest FG: Common occurrence of enterotoxin genes and enterotoxicity in Bacillus thuringiensis. FEMS Microbiol Lett, 190, 151-155, 2000.
33. Hansen BM, Hendriksen NB: Detection of enterotoxic Bacillus cereus and Bacillus thuringiensis strains by PCR analysis. Appl Environ Microbiol, 67, 185-189, 2001.
34. Zahner V, Cabral DA, Régua-Mangia AH, Rabinovitch L, Moreau G, McIntosh D: Distribution of genes encoding putative virulence factors and fragment length polymorphisms in the vrrA gene among Brazilian isolates of Bacillus cereus and Bacillus thuringiensis. Appl Environ Microbiol, 71, 8107-8114, 2005.
35. Mäntynen V, Lindström K: A rapid PCR-based DNA test for enterotoxic Bacillus cereus. Appl Environ Microbiol, 64, 1634-1639, 1998.
36. Nduhiu GJ, Jackson NO, Dorcas WN: Genetic characterisation of food borne Bacillus cereus strains from milk, cheese and rice by multiplex PCR assay. Int J Integr Biol, 5, 82-86, 2009.
37. Beecher DJ: The Bacillus cereus group. In, Sussman M (Ed): Molecular Medical Microbiology. 1161-1190, Academic Press, New York, 2001.
38. Guinebretiére MH, Broussolle V, Nguyen-The C: Enterotoxigenic profiles of food-poisoning and food-borne Bacillus cereus strains. Appl Environ Microbiol, 40, 3053-3056, 2002.
39. Moravek M, Wegscheider M, Schulz A, Dietrich R, Bürk C, Märtlbauer E: Colony immunoblot assay for the detection of hemolysin BL enterotoxin producing Bacillus cereus. FEMS Microbiol Lett, 238, 107-113, 2004.
40. Yang IC, Shih DY, Huang TP, Huang YP, Wang JY, Pan TM: Establishment of a novel multiplex PCR assay and detection of toxigenic strains of the species in the Bacillus cereus group. J Food Protect, 68, 2123-2130, 2005.
41. Ngamwongsatit P, Buasri W, Pianariyanon P, Pulsrikam C, Ohba M, Assavanig A, Panbangred W: Broad distribution of enterotoxin genes (hblCDA, nheABC, cytK, and entFM) among Bacillus thuringiensis and Bacillus cereus as shown by novel primers. Int J Food Microbiol, 121, 352356, 2008.
42. Lund T, De Buyser ML, Granum PE: A new cytotoxin from Bacillus cereus that may cause necrotic enteritis. Mol Microbiol, 38, 254-261, 2000.

APA	ÇADIRCI Ö, gücükoğlu a, TERZİ GÜLEL G, KEVENK T, ALİŞARLI M (2013). Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR. , 869 - 874.
Chicago	ÇADIRCI Özgür,gücükoğlu ali,TERZİ GÜLEL GÖKNUR,KEVENK Tahsin Onur,ALİŞARLI MUSTAFA Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR. (2013): 869 - 874.
MLA	ÇADIRCI Özgür,gücükoğlu ali,TERZİ GÜLEL GÖKNUR,KEVENK Tahsin Onur,ALİŞARLI MUSTAFA Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR. , 2013, ss.869 - 874.
AMA	ÇADIRCI Ö,gücükoğlu a,TERZİ GÜLEL G,KEVENK T,ALİŞARLI M Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR. . 2013; 869 - 874.
Vancouver	ÇADIRCI Ö,gücükoğlu a,TERZİ GÜLEL G,KEVENK T,ALİŞARLI M Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR. . 2013; 869 - 874.
IEEE	ÇADIRCI Ö,gücükoğlu a,TERZİ GÜLEL G,KEVENK T,ALİŞARLI M "Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR." , ss.869 - 874, 2013.
ISNAD	ÇADIRCI, Özgür vd. "Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR". (2013), 869-874.

APA	ÇADIRCI Ö, gücükoğlu a, TERZİ GÜLEL G, KEVENK T, ALİŞARLI M (2013). Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 19(5), 869 - 874.
Chicago	ÇADIRCI Özgür,gücükoğlu ali,TERZİ GÜLEL GÖKNUR,KEVENK Tahsin Onur,ALİŞARLI MUSTAFA Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 19, no.5 (2013): 869 - 874.
MLA	ÇADIRCI Özgür,gücükoğlu ali,TERZİ GÜLEL GÖKNUR,KEVENK Tahsin Onur,ALİŞARLI MUSTAFA Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, vol.19, no.5, 2013, ss.869 - 874.
AMA	ÇADIRCI Ö,gücükoğlu a,TERZİ GÜLEL G,KEVENK T,ALİŞARLI M Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR. Kafkas Üniversitesi Veteriner Fakültesi Dergisi. 2013; 19(5): 869 - 874.
Vancouver	ÇADIRCI Ö,gücükoğlu a,TERZİ GÜLEL G,KEVENK T,ALİŞARLI M Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR. Kafkas Üniversitesi Veteriner Fakültesi Dergisi. 2013; 19(5): 869 - 874.
IEEE	ÇADIRCI Ö,gücükoğlu a,TERZİ GÜLEL G,KEVENK T,ALİŞARLI M "Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR." Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 19, ss.869 - 874, 2013.
ISNAD	ÇADIRCI, Özgür vd. "Determination of enterotoxigenic gene profiles of bacillus cereus strains isolated from dairy desserts by multiplex PCR". Kafkas Üniversitesi Veteriner Fakültesi Dergisi 19/5 (2013), 869-874.