Investigation of Toxin-Producing and Antibiotic-Resistant Bacillus cereus in Spices Used in the Production of Sucuk

Year 2021, , 331 - 340, 31.12.2021

Ece Çetin , Görkem Yaman , Artun Yıbar

https://doi.org/10.31590/ejosat.1010007

Abstract

In this study, a total of 100 samples of seven types of spices were investigated to determine the presence of toxin-producing and multiple antibiotic-resistant Bacillus cereus and other food-borne microorganisms. The spices tested included 18 samples of ground black pepper (Piper nigrum), 17 samples of red pepper (Capsicum frutescens), 12 samples of ground red pepper (Capsicum frutescens), 20 samples of cummin (Cuminum cyminum), 16 samples of pimento (Pimenta dioica), nine samples of garlic powder (Allium sativum) and eight samples of sucuk spice mixes that were obtained from various retail shops and sucuk production units in the Bursa province between January and December 2014. These spices are used to prepare sucuk because of their flavouring and seasoning properties. The samples were analysed for Bacillus cereus, Escherichia coli, coagulase-positive Staphylococcus aureus, Salmonella spp., yeast, moulds and coliforms. For the identification of B. cereus, Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS, Germany) was used. The analysis showed that the samples had coliform, yeast and mould and B. cereus counts between <10 and 1.1x105 cfu/g, 1x102 and 9.3x104 cfu/g, and 1x102 and 4.8x104 cfu/g, respectively. Furthermore, coagulase-positive S. aureus was identified in five samples, E. coli was identified in two samples, and Salmonella spp. was identified in none. A total of 39 (92.8%) B. cereus isolates were resistant to two or more antibiotic agents. In addition, 31 (73.8%) B. cereus isolates tested positive for production of both non-haemolytic enterotoxin (NHE) and haemolysin BL (HBL) enterotoxins. These results show that some spices contain harmful microorganisms. To reduce microbial counts and to improve poor microbiological quality in spices, hygiene programmes must be corrected in the pre- and post-harvesting period.

Keywords

spices, Sucuk, Bacillus cereus, Multiple antibiotic resistance, Toxin producing

Thanks

This manuscript was edited before submission by American Journal Experts (AJE).

Sucuk'un Üretiminde Kullanılan Baharatlarda Toksin Üreten ve Antibiyotiğe Dirençli Bacillus cereus'un Araştırılması

Abstract

Bu çalışmada, toksin üreten ve çoklu antibiyotiklere dirençli Bacillus cereus ve diğer gıda kaynaklı mikroorganizmaların varlığını belirlemek için yedi çeşit baharattan toplam 100 numune araştırılmıştır. Çalışmanın amacı doğrultusunda; Ocak ve Aralık 2014 tarihleri arasında Bursa ilinde, çeşitli perakende satış mağazaları ve sucuk üretim ünitelerinden elde edilen 18 kara biber örneği (Piper nigrum), 17 kırmızı biber örneği (Capsicum frutescens), 12 toz kırmızı biber örneği (Capsicum frutescens), 20 kimyon numunesi (Cuminum cyminum), 16 yenibahar örneği (Pimenta dioica), dokuz sarımsak tozu (Allium sativum) ve sekiz sucuk baharatı karışımları örnekleri analiz edilmiştir. Bu baharatlar, aromaları ve baharat özelliklerinden dolayı sucuk hazırlamak için kullanılmaktadır. Örnekler, Bacillus cereus, Escherichia coli, koagülaz pozitif Staphylococcus aureus, Salmonella spp., Maya-küf ve koliformlar yönünden analiz edildmiştir. Bacillus cereus’un tanımlanmasında Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS, Germany) kullanıldı. Analizler, örneklerin koliform, maya ve küf ve B. cereus sayımlarının sırasıyla <10 and 1.1x105 cfu/g, 1x102 and 9.3x104 cfu/g, and 1x102 and 4.8x104 cfu/g arasında olduğunu gösterdi. Ayrıca, beş örnekte koagülaz pozitif S. aureus, iki örnekte E. coli belirlenirken, hiçbir örnekte Salmonella spp. varlığı tespit edilemedi. Toplam 39 (% 92.8) B. cereus izolatının iki veya daha fazla antibiyotik ajana dirençli olduğu belirlendi. Ek olarak, B. cereus izolatlarının 31 (% 73.8) adeti non-hemolitik enterotoksin (NHE) ve hemolizin BL (HBL) enterotoksinlerinin üretimi için pozitif test edildi. Bu sonuçlar, bazı baharatların zararlı mikroorganizmaları içerdiğini göstermektedir. Mikrobiyal sayımı azaltmak ve baharatlarda zayıf mikrobiyolojik kaliteyi arttırmak için hijyen programları hasat öncesi ve hasat sonrası düzeltilmelidir.

Keywords

Baharat, Sucuk, Bacillus cereus, Çoklu antibiyotik direnci, Toksin üretimi, MALDİ-TOF

References

• Agaoglu, S., Sancak, Y. C., Alisarli, M., Ekici, K. 1999. The presence of B. cereus in some spice varieties offered for sale in Van market. Journal of Research in Veterinary Medicine, 1-2(18), 89-96.
• Aksu, H., Bostan, K., Ergün, Ö. 2000. Presence of Bacillus cereus in packaged some spices and herbs sold in Istanbul. Pakistan Journal of Biological Sciences, 3(5), 710-712.
• Antai, S. P., 1988. Study of Bacillus flora of Nigerian spices. International Journal of Food Microbiology, 6: 259-261.
• Banerjee, M., Sarkar, P. K. 2003. Microbiological quality of some retail spices in India. Food Research International, 36: 469-474.
• Bauer, A. W., Kirby, W. M., Sherris, J. C., Truck, M. 1966. Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology, 45: 493-496.
• Brown, J. C., Jiang, X. 2008. Prevalence of antibiotic-resistant bacteria in herbal products. Journal of Food Protection, 71(7), 1486-90.
• Cameron, G. 1998. Need to consider Bacillus subtilis as a cause of food poisoning. New Zealand Public Health Surveillance Report, 5(2), 11.
• Deambrosis, N., Da Silva, A. 1992. Incidence of Bacillus cereus in spices. Proceedings of the 3rd World Congress on Feodborne Infections and Intoxications, 1: 316, Berlin.
• De Boer, E., Spielenberg, W. M., Janssen, F. W. 1985. Microbiology of spice and herbs. Antonie Van Leeuwenhoek International Journal of General and Molecular Micr, 51: 435-438.
• Debs-Louka, E., Zouki, J. E., Dabboussi, F. 2013. Assessment of the microbiological quality and safety of common spices and herbs Sold in Lebanon, Journal of Food and Nutritional Disorders, 2: 4.
• Eglezos, S., Huang, B., Dykes, E. A., Fegan, N. 2010. The prevalence and concentration of Bacillus cereus in retail food products in Brisbane, Australia. Foodborne Pathogens and Disease, 7 (7), 1-3.
• Elmalı, M., Yaman, H. 2005. Microbiological Quality of some spices sold in the Markets of Bitlis District. Erciyes University Journal of the Faculty of Veterinary Medicine, 2(1), 9-14.
• Erol, İ., Küplülü, Ö., Karagöz, S. 1999. Microbiological quality of some spices consumed in Ankara. Veterinary Journal of Ankara University, 46: 115-125.
• Filiz, N. 2001. Microbial flora of some ground spices consumed in Bursa, Journal of Research in Veterinary Medicine, 20: 103-107.
• García, S., Iracheta, F., Galván, F., Heredia, N. 2001. Microbiological survey of retail herbs and spices from Mexican markets. Journal of Food Protection, 64: 99-103.
• Giaccone, V., Colavita, G., Torriani, S., Ciocca, R. M., Augelli, R. 1996. Occurrence of Bacillus cereus and other Bacillus spp. in spices. Archiv Für Lebensmittelhygiene, 47(2), 47-49.
• Guinebretiere, M. H., Broussolle, V., Nguyen-The, C. 2002. Enterotoxigenic profiles of food-poisoning and food-borne Bacillus cereus strains. Journal of Clinical Microbiology, 40: 3053-3056.
• Hampikyan, H., Bingol, E. B., Colak, H., Aydin, A. 2009. The evaluation of microbiological profile of some spices used in Turkish meat industry. Journal of Food, Agriculture and Environment, 7(3-4), 111-115.
• Hansen, B. M., Hendriksen, N. B.2001. Detection of enterotoxic Bacillus cereus and Bacillus thuringiensis strains by PCR analysis. Applied and Environmental Microbiology Journal, 67(1), 185-189. https://doi.org/10.1128/AEM.67.1.185-189.2001.
• Hariram, U., Labbé, R.2015. Spore prevalence and toxigenicity of Bacillus cereus and Bacillus thuringiensis isolates from U.S. retail spices. Journal of Food Protection, 78(3), 590-596.
• Hassan, S. A., Altalhi, A. D. 2013. Safety assessment of spices and herbs consumed in Saudi Arabia: Microbiological Quality and Toxin Production. Life Science Journal, 10(4), 2819-2827.
• International Organization for Standardization (ISO 6579: 2002). 2002. Microbiolgy of food and animal feeding stuffs-Horizontal method for detection of Salmonella spp. Geneva, Switerland.
• Julseth, R. M., Deibel, R. H. 1974. Microbial profile of selected spices and herbs at Import. Journal of Milk and Food Technology, 37: 414-419.
• Khachatourians, G. G. 1998. Agricultural use of antibiotics and the evolution and transfer of antibiotic-resistant bacteria. Canadian Medical Association Journal, 159(9), 1129-1136.
• Kneifel, W., Berger, E. 1994. Microbiological criteria of random samples of spices and herbs retailed on the Austrian Market. Journal of Food Protection, 57: 839-901.
• Konuma, H., Shinagawa, K., Tokumaru, M., Onoue, Y., Konno, S., Fujino, N., Shigehisa, T., Kurata, H., Kuwabara, Y., Lopes, C. A. M. 1988. Occurrence of Bacillus cereus in meat products, raw meat and meat product additives. Journal of Food Protection, 51(4), 324-326.
• Koohy-Kamaly-Dehkordy, P., Nikoopour, H., Siavoshi, F., Koushki, M., Abadi, A. 2013. Microbiological quality of retail spices in Tehran, Iran. Journal of Food Protection, 76(5), 843-852. https://doi.org/10.4315/0362-028X.JFP-12-180
• Kramer, J. M., Gilbert, R. J. 1989. Bacillus cereus and other Bacillus species. In, Doyle MP (Ed): Foodborne Bacterial Pathogens. 21-70, Marcel Dekker, New York.
• Kovács-Domján, H. 1988. Microbioligical investigation of paprika and pepper with special regard to sporeformers including B. cereus. Acta Alimentaria, 17: 257-264.
• Little, C. L., Omotoye, R., Mitchell, R. T. 2003. The microbiological examination of ready-to-eat foods to which spices have been added. International Journal of Environmental Health Research, 13(1), 31-42.
• Lund, T., Granum, P. E. 1996. Characterisation of a non-haemolytic enterotoxin complex from Bacillus cereus isolated after a foodborne outbreak. FEMS Microbiology Letters, 141(2-3), 151-6.
• Meena, B. S., Kapoor, K. N., Agarwal, R. K. 2000. Occurrence of multi-drug resistant Bacillus cereus in foods. Journal of Food Science and Technology, 37: 289-291.
• Moravek, M., Dietrich, R., Buerk, C., Broussolle, V., Guinebretière, M. H., Granum, P. E., Nguyen-The, C., Märtlbauer, E. 2006. Determination of the toxic potential of Bacillus cereus isolates by quantitative enterotoxin analyses. FEMS Microbiology Letters, 257(2), 293-8.
• Moss, M. O., Baker, T. 2000. Teaching Techniques for Mycology: 10. Moulds from spices; Eurotium species. Mycologist, 14(2), 54-57.
• Pafumi, J. 1986. Assessment of the microbiological quality of spices and herbs. Journal of Food Protection, 49(12), 958-963.
• Parveen, S., Das, S., Begum, A., Sultana, N., Hoque, M. M., Ahmad, I. 2014. Microbiological quality assessment of three selected spices in Bangladesh. International Food Research Journal, 21(4), 1327-1330.
• Phillips, I., Casewell, M., Cox, T., Brad De Groot, Friis, C., Jones, R., Nightingale, C., Preston, R., Waddell, J. 2004. Does the use of antibiotics in food animals pose a risk to human health? A critical review of published data. Journal of Antimicrobial Chemotherapy, 53: 28-52. https://doi.org/10.1093/jac/dkg483
• Rosenberger, A., Weber, H. 1993. Keimbelastung von gewurzproben. Mikrobiologischer status im hinblick auf richt und warnwerte. Fleischwirtsch, 73: 830-833.
• Powers, E. M., Latt, T. G., Brown, T. 1976. Incidence and levels of Bacillus cereus in processed spices. Journal of Milk and Food Technology, 39(10), 668-670.
• Sagoo, S. K., Little, C. L., Greenwood, M., Mithani, V., Grant, K. A., McLauchlin, J., Pinna, E., Threlfall, E. J. 2009. Assessment of the microbiological safety of dried spices and herbs from production and retail premises in the United Kingdom. Food Microbiology, 26: 39-43.
• Schoenı, J. L., Wong, A. C. L. 2005. Bacillus cereus food poisoning and its toxins. Journal of Food Protection, 68(3), 636-648.
• Shinagawa, H., Makino, K., Amemura, M., Kimura, S., Iwasaki, H., Nakata, A. 1988. Structure and regulation of the Escherichia coli ruv operon involved in DNA repair and recombination. Journal of Bacteriology, 170: 4322-4329.
• Te Giffel, M. C., Beumer, R. P., Granum pe, Rombouts, F. M.1997. Isolation and characterisation of Bacillus cereus from pasteurised milk in household refrigerators in the Netherlands. International Journal of Food Microbiology, 34, 307-318.
• Temelli, S., Anar, S. 2002. Prevalence of Bacillus cereus in spices and herbs marketed in Bursa. Journal of Faculty of Veterinary Medicine, Istanbul University, 28(2), 459-465.
• Tewari, A., Singh, S. P., Singh, R. 2012. Prevalence of multidrug resistant Bacillus cereus in foods and human stool samples in and around Pantnagar, Uttrakhand. Journal of Advanced Veterinary and Animal Research, 2: 252-255.
• Turkısh Food Codex Mıcrobıologıcal Crıterıa Regulatıon, 2011.
• Third Trimester National Microbiological Survey, 2005. EU Coordinated programme. Bacteriological Safety of Pre-packaged Mixed Salads.
• Tulu S, Demissie S, Dabassa A: Mikrobial safety of some selected spices sold in Jimma Town, South Western of Ethiopia. Res J Microbiol, 9(1): 43-50, 2013.
• Verluyten, J., Leroy, F., Luc de Vuyst. 2004. Effects of different spices used in production of fermented sausages on growth of and curvacin a production by Lactobacillus curvatus LTH 1174. Applied and Environmental Microbiology Journal, 70(8), 4807-4813. https://doi.org/10.1128/AEM.70.8.4807-4813.2004
• Vural, A. 2004. Investigation on the microbiological quality of consumed some spices in Diyarbakır province. Kafkas Universitesi Veteriner Fakultesi Dergisi, 10(1), 13-18.
• Wehrle, E., Moravek, M., Dietrich, R., Bürk, C., Didier, A., Märtlbauer, E. 2009. Comparison of multiplex PCR, enzyme immunoassay and cell culture methods for the detection of enterotoxinogenic Bacillus cereus. Journal of Microbiological Methods, 78: 265-270.
• Whong, C. M. Z., Kwaga, J. K. P. 2007. Antibiograms of Bacillus cereus isolates from some Nigerian foods. Nigerian Food Journal, 25(1), 178-183.
• Yang, I. C., Shih, D. Y., Huang, T. P. 2005. Establishment of a novel multiplex PCR assay and detection of toxigenic strains of the species in the Bacillus cereus group. Journal of Food Protection, 68: 2123-30. https://doi.org/10.4315/0362-028X-68.10.2123