Didem ÖZER, Ayten ERDEM KİMİRAN

Comparison of different methods for the detection of salmonella spp. in minced meat samples

İstanbulun dokuz farklı semtinde tüketime sunulan kıyma örneklerinde geleneksel kültür yöntemi (ISO), immünomanyetik ayırma (İMA) ve floresanlı yerinde hibritleme (FISH) yöntemleri kullanılarak Salmonella cinsi bakterilerin varlığı tespit edilmiştir. İncelenen 50 kıyma örneğinin, geleneksel kültür yöntemi ile 5 inden (20%), İMA yöntemi ile 3 ünden (6%) Salmonella cinsi bakteri izole edilmiştir. Gerek geleneksel kültür yöntemi gerekse de İMA yönteminde besiyeri üzerinde üreyen kolonilerden elde edilen izolatlar Salmonella choleraesuis ssp. arizonae ve Salmonella spp. olarak tanımlanmıştır. İncelenen 50 kıyma örneğinin 37sinde (74%) ise FISH yöntemi ile Salmonella cinsi bakterilerin varlığı tespit edilmiştir. Çalışmamızda Salmonella cinsi bakterilerin varlığını belirlemede geleneksel kültür yönteminin, İMA yöntemine göre daha duyarlı bir yöntem olduğu, FISH yönteminin ise Salmonella cinsi bakterilerin varlığını belirlemede en iyi yöntem olduğu tespit edilmiştir. Gıda kaynaklı patojenlerle kontaminasyon sonucunda ortaya çıkan uluslararası önemdeki salgınların hızlı tespiti ne kadar önemli olsa da, uygulanan yeni yöntemlerin sonuçlarının geleneksel kültür yöntemi ile desteklenmesi gerekmektedir.

Kıyma örneklerinde salmonella spp. tespitinde farklı yöntemlerin karşılaştırılması

The presence of Salmonella spp. in minced meat that is consumed in nine different sites of Istanbul is evaluated by using conventional culture (ISO 6579:2002), immunomagnetic separation (IMS) and fluorescent in situ hybridization (FISH) methods. Salmonella spp. was isolated from five of 50 (20%) minced meat with ISO 6579 method, and three of 50 (6%) minced meat using IMS method. Bacteria isolated from both ISO and IMS methods were identified as Salmonella choleraesuis ssp. arizonae and Salmonella spp. The presence of Salmonella spp. was determined from 37 of 50 (74%) minced meat by using FISH method. In the current study, it has been shown that ISO 6579 method was found more to be susceptible than IMS method for determining presence of Salmonella spp., FISH method is the best method to determine the presence of Salmonella spp. Even if the quick determination of the epidemics of international importance occured as a result of the contamination by pathogens derivated from foods, the results of the use of new methods should be supported by the conventional culture method.

PDF

___

1. Farkas J: Irradiation as a method for decontaminating food. Int J Food Microbiol, 44, 189-204, 1998.
2. Sekin Y, Karagözlü N: Gıda Mikrobiyolojisi. Gıda Endüstrisi İçin Temel Esaslar ve Uygulamalar. Literatür Yayıncılık, İstanbul, 2004.
3. Güven A, Gülmez M, Kamber U: Kars ilinde tüketime sunulan kıymalarda bazı patojen mikroorganizmaların araştırılması ve kıymaların mikrobiyolojik kalitesinin belirlenmesi. Kafkas Univ Vet Fak Derg, 3 (1): 57 65, 1997.
4. Jeníková G, Pazlarová J, Demnerová K: Detection of Salmonella in food samples by the combination of immunomagnetic separation and PCR assay. Int Microbiol, 3, 225-229, 2000.
5. Erol I: Ankarada tüketime sunulan kıymalarda Salmonellaların varlığı ve serotip dağılımı. Turk J Vet Anim Sci, 23, 321-325, 1997.
6. Favrin SJ, Jassim SA, Griffiths MW: Application of a novel immuno magnetic separation-bacteriophage assay for the detection of Salmonella enteritidis and Escherichia coli O157:H7 in food. Int J Food Microbiol, 85, 63-71, 2003.
7. Pyle BH, Broadaway SC, Mcfeters GA: Sensitive detection of Escherichia coli O157:H7 in food and water by immunomagnetic separation and solid-phase laser cytometry. Appl Environ Microbiol, 65 (5): 1966-1972, 1999.
8. Mansfield LP, Forsythe SJ: Immunomagnetic separation as an alternative to enrichment broths for Salmonella detection. Lett Appl Microbiol, 16, 122-125, 1993.
9. Skjerve E, Rorvik LM, Olsvik O: Detection of Listeria monocytogenes in foods using immunomagnetic separation. Appl Environ Microbiol, 56, 3478-3481, 1990.
10. Skjerve E, Olsvik O: Immunomagnetic separation of Salmonella from foods. Int J Food Microbiol, 14, 11-18, 1991.
11. Sonti SV, Bose A: Cell separation using protein A coated magnetic nanoclusters. J Colloid Interface Sci, 170, 575-585, 1995.
12. Coleman DJ, Chick KE, Nye KJ: An evaluation of immunomagnetic separation for the detection of Salmonella s in raw chicken carcasses. Lett Appl Microbiol, 21, 152-154, 1995.
13. Cudjoe KS, Krona R: Detection of Salmonella from raw food samples using Dynabeads anti-Salmonella and a conventional reference method. Int J Food Microbiol, 37 (1): 55-62, 1997.
14. Rijpens N, Herman L, Vereecken F, Jannes G, De Smedt J, De Zutter L: Rapid detection of stressed Salmonella spp. in dairy and egg products using immunomagnetic separation and PCR. Int J Food Microbiol, 46, 37-44, 1999.
15. Cocolin L, Diez A, Urso R, Rantsiou K, Comi G, Bergmaier I, Beimfohr C: Optimization of conditions for profiling bacterial populations in food by culture - independent methods. Int J Food Microbiol, 120 (1-2): 100-109, 2007.
16. Ripabelli G, Sammarco ML, Ruberto A, Iannitto G, Grasso GM: Immuno-magnetic separation and conventional culture procedure for detection of naturally occurring Salmonella in raw pork sausages and chicken meat. Lett Appl Microbiol, 24, 493-497, 1997.
17. Vieira - Pinto MV, Oliveira M, Aranha J, Martins C, Bernardo F: Influence of an enrichment step on Salmonella spp. detection by fluorescent in situ hybridization on pork samples. Food Control, 19, 286- 290, 2008.
18. Mansfield LP, Forsythe SJ: The detection of Salmonella serovars from animal feed and raw chicken using a combined immunomagnetic separation and ELISA method. Food Microbiol, 18 (4): 361-366, 2001.
19. Taban B: İmmünomanyetik ayırma-polimeraz zincir reaksiyonu yönte-minin uygulanması ile tavuk etlerinde Salmonella spp. belirlenmesi. Doktora tezi, Hacettepe Üniv. Fen Bil. Enst., 2007.
20. Mercanoglu B, Aytac SA, Ergun MA, Tan E: Isolation of Listeria monocytogenes by immunomagnetic separation and atomic force micro-scopy. J Microbiol, 41 (2): 144-147, 2003.
21. Riberio AR, Nascimento VP, Cardoso MO, Santos LR, Rocha SL: Utilization of immunomagnetic separation for detection of Salmonella in raw broiler parts. Braz J Microbiol, 33 (4): 339-341, 2002.
22. Boer E, Beumer RR: Methodology for detection and typing of foodborne microorganisms. Int J Food Microbiol, 50, 119-130, 1999.
23. Tekinsen OC, Yurtyeri A, Mutluer B: Ankara`da satılan hazır kıymaların bakteriyolojik kalitesi. Ankara Univ Vet Fak Derg, 27 (1-2): 45-63, 1980.
24. Reddy SG, Henrickson RL, Olson HC: The influence of lactic cultures on ground beef quality. J Food Sci, 35, 787-781, 1970.
25. Sarıgöl C: Elazığda tüketilen kıymalarda Clostridium ve Enterobacteriaceae grubu mikroorganizmaların varlığı üzerinde araştırmalar. Fırat Univ Vet Fak Derg, 7, 179-186, 1982.
26. Gökalp HY, Yetim H, Karacam H: Some saprophytic and pathogenic bacteria levels of ground beef sold in Erzurum, Turkey. In, Proceeding of 2. World Congress of Foodborne Infections and Intoxications, Berlin, Germany, pp. 310-313, 1982.
27. Aabo S, Andersen JK, Olsen JE: Detection of Salmonella in minced meat by the polymerase chain reaction method. Lett Appl Microbiol, 21, 180-182, 1995.
28. Straub TM, Dockendorff BP, Quiñonez-Díaz MD, Valdez CO, Shutthanandan JI, Tarasevich BJ, Grate JW, Bruckner-Lea CJ: Automated methods for multiplex pathogen detection. J Microbiol Methods, 62, 303-316, 2005.
29. Coleman DJ, Nye KJ, Chick KE, Gagg CM: A comparison of immuno magnetic separation plus enrichment with conventional Salmonella culture in the examination of raw sausages. Lett Appl Microbiol, 21, 249 -251, 1995.
30. Molla B, Kleer J, Sinell HJ: Detection of Salmonella in foods by immuno-magnetic separation. Archiv für Lebensmittelhyg, 45 (5): 110-113, 1994.
31. Blackburn W, De C, Patel PD, Gibbs PA: Separation and detection of Salmonella using immunomagnetic particles. Biofouling, 5, 143-155, 1991.
32. Okrend AJG, Rose BE, Lattuada CP: Isolation of Escherichia coli O157:H7 using O157 specific antibody coated magnetic beads. J Food Protect, 55, 214-217, 1992.
33. Vermunt AEM, Franken AAJM, Beumer RR: Isolation of Salmonellas by immunomagnetic separation. J Appl Bacteriol, 72, 112-118, 1992.
34. Fratamico PM: Comparison of culture, polymerase chain reaction (PCR), TaqMan Salmonella, and transia card Salmonella assays for detection of Salmonella spp. in naturally-contaminated ground chicken, ground turkey and ground beef. Mol Cell Probes, 17, 215-221, 2003.
35. Gökmen M, Alişarlı M: Van ilinde tüketime sunulan kıymaların bazı patojen bakteriler yönünden incelenmesi. YYU Vet Fak Derg, 14 (1): 27-34, 2003.
36. Vieira - Pinto M, Olivieira M, Bernardo F, Maruns C: Evaluation of fluorescent in situ hybridization (FISH) as a rapid screening method for detection of Salmonella in tonsils of slaughtered pigs for consumption: A comparison with conventional culture method. J Food Safety, 25 (2): 109-119, 2005.
37. Blasco L, Ferrer S, Pardo I: Development of specific fluorescent oligonucleotide probes for in situ identification of wine lactic acid bacteria. FEMS Microbiol Lett, 225, 115-123, 2003.
38. Amann RI, Ludwig W, Schleifer KH: Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Am Soc Microbiol, 59 (1): 143-169, 1995.
39. Niederhauser C, Candrian U, Hofelein C, Jermini M, Buhler HP, Luthy J: Use of polymerase chain reaction for detection of Listeria monocytogenes in food. Appl Environ Microbiol, 58 (5): 1564-1568, 1992.
40. Stender H, Oliveira K, Rigby S, Bargoot F, Coull J: Rapid detection, identification, and enumeration of Escherichia coli by fluorescence in situ hybridization using an array scanner. J Microbiol Methods, 45, 31-39, 2001.