Clostridium botulinum Spores Found in Honey from Small Apiaries in Poland

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Abstract

A total of 102 honey samples collected from small apiaries (≤ 20 hives) in Poland were analysed for the presence of Clostridium botulinum spores. The samples were prepared using the dilution centrifugation method and cultured in parallel in cooked meat medium (CMM) and tripticase peptone glucose yeast (TPGY) enrichment broths. Identification of toxin types A, B, and E of Clostridium botulinum strains was performed with the use of the multiplex PCR method. Positive samples were also subjected to quantitative analysis with the use of Clostridium botulinum Isolation Agar Base (CBAB). The prevalence analysis showed 22 (21.6%) samples contaminated with C. botulinum spores. The major serotype detected was botulin neurotoxin type A – 16 (72.7%) whereas type B was found in 3 (13.6%) honey samples and type E also only in 3 (13.6%) honey samples. Dual-toxin-producing strains were noted. The average quantity of spores in PCR - C. botulinum positive samples was 190 in 1 gram of honey.

Arnon, S.S., Midura, T.F., Damus, K., Thompson, B., Wood, R.M., Chin, J. (1979). Honey and other environmental risk factors for infant botulism. The Journal of Pediatrics, 94(2), 331–336. http://doi.org/10.1016/S0022-3476(79)80863-X

Arnon, S.S. (1998). Infant botulism. In Textbook of pediatric infectious diseases. (pp. 1758-1766). Philadelphia: Elsevier Inc.

Arnon, S.S., Schechter, R., Inglesby, T.V., Henderson, D.A., Bartlett, J.G., Ascher, M.S., ... Tonat, K. (2001). Botulinum toxin as a biological weapon: medical and public health management. The Journal of the American Medical Association, 285(8), 1059-1070. http://doi.org/10.1001/jama.285.8.1059

Aureli, P., Franciosa, G., & Fenicia, L. (2002). Infant botulism and honey in Europe: a commentary. The Pediatric Infectious Disease Journal, 21(9), 866-868. http://doi.org/10.1097/01.inf.0000027418.41135.3f

Babapour, A., Razavilar, V., Arzanlou, M., Khazdouzm, A. (2015). Study on prevalence of humen toxogenic types (A, B, E) of C. botulinum by nested PCR method in domestic and commercial honeys in Ardebil (Northwest of Iran). International Journal of Biology, Pharmacy and Allied Sciences, 4(2), 334–346.

Bianco, M.I., Luquez, C., de Jong, L.I.T., Fernanadez, R.A. (2009). Linden flower (Tilia spp.) as potential vehicle of Clostridium botulinum spores in the transmission of infant botulism. Revista Argentina de Microbiologia, 41(4), 232-236.

Brook, I. (2007). Infant botulism. Journal of Perinatology, 27, 175–180. http://doi.org/10.1038/sj.jp.7211651

Ciećko, Z., Wyszkowski, M., Krajewski, W., Zabielska J. (2001). Effect of organic matter and liming on the reduction of cadmium uptake from soil by triticale and spring oilseed rape. Science of The Total Environment, 2819(1–3), 37–45. http://doi.org/10.1016/S0048-9697(01)00800-2

Critchley, E.M. (1991). A comparison of human and animal botulism: a review. Journal Of the Royal Society of Medicine, 84(5), 295–298. http://doi.org/10.1177/014107689108400517

Dahlenborg, M., Borch, E. & Radstro, P. (2001). Development of a combined selection and enrichment PCR procedure for Clostridium botulinum types B, E, and F and its use to determine prevalence in fecal samples from slaughtered pigs. Applied and Environmental Microbiology, 67(10), 4781–4788. http://doi.org/10.1128/AEM.67.10.4781–4788.2001

Estevinho, L.M., Feás, X., Seijas, J.A., Vazquez-Tato M.P. (2012). Organic honey from Trás-Os-Montes region (Portugal): Chemical, palynological, microbiological and bioactive compounds characterization. Food and Chemical Toxicology, 50(2), 258-264. http://doi.org/10.1016/j.fct.2011.10.034

Gale, P., Drew, T., Phipps, L.P., David, G., Wooldridge M. (2009). The effect of climate change on the occurrence and prevalence of livestock diseases in Great Britain: a review. Journal of Applied Microbiology, 106(5), 1409-1423. http://doi.org/10.1111/j.1365-2672.2008.04036.x

Gilbert, S., Lake, R., Hudson, A., Cressey P. (2006). Risk profile: C. botulinum in honey. Institute of Environmental Science & Research Limited. Christchurch Science, Centre New Zeeland. Retrieved May 1, 2016, from http://www.foodsafety.govt.nz/elibrary/industry/Risk_Profile_Clostridium-Science_Research.pdf

Horowitz, B.Z. (2005). Botulinum toxin. Critical Care Clinics, 21(4), 825–839. http://doi.org/10.1016/j.ccc.2005.06.008

Huhtanen, C.N., Knox, D. & Shimanuki H. (1981). Incidence and origin of Clostridium botulinum spores in honey. Journal of Food Protection, 44(11), 812-814

Huss, H.H. (1980). Distribution of Clostridium botulinum. Applied and Environmental Microbiology, 39(4), 764-769.

Johannsen, A. (1963). Clostridium botulinum in Sweden and the adjacent waters. Journal of Applied Bacteriology, 26(1):43-7.

Kędzia, B., & Hołderna-Kędzia, E. (2008). Miód. Skład i właściwości biologiczne. Warszawa: PW „Rzeczpospolita” SA.

Kędzia, B., & Hołderna-Kędzia, E. (2010). Lecznicze działanie miodu pszczelego w chorobach wewnętrznych. Wrocław: MedPharm Polska.

Koluman, A., Melikoglu Golcu, B., Derin, O., Ozkok, S., Anniballi F. (2013). Clostridium botulinum in honey: prevalence and antibiotic susceptibility of isolated strains. Turkish Journal of Veterinary and Animal Sciences, 37, 706-711. http://doi.org/10.3906/vet-1209-40

Küplülü, Ö., Göncüoğlu, M., Özdemir, H., Koluman A. (2006). Incidence of Clostridium botulinum spores in honey in Turkey. Food Control, 17(3), 222-224. http://doi.org/10.1016/j.foodcont.2004.10.004

Lindström, M., & Korkeala, H. (2006). Laboratory diagnostics of botulism. Clinical Microbiology Reviews, 19(2), 298-314.http://10.1128/CMR.19.2.298-314.2006

Loncaric, I., Ruppitsch, W., Licek, E., Moosbeckhofer, R., Busse, H.J., Rosengarten, R. (2011). Characterization of selected Gram-negative non-fermenting bacteria isolated from honey bees (Apis melifera carnica). Apidologie, 42(3), 312-325. http://doi.org/10.1007/s13592-011-0019-7

Long, S.S. (2001). Infant botulism. The Pediatric Infectious Disease Journal, 20(7), 707–709.

Luquez, C., Bianco, M.I., de Jong, L.I.T., Sagua, M.D., Arenas, G.N., Ciccarelli, A.S., Fernandez, R.A. (2005). Distribution of botulinum toxin-producing Clostridia in soils of Argentina. Applied and Environmental Microbiology, 71(7), 4137-4139. http://doi.org/10.1128/AEM.71.7.4137-4139.2005

Madras-Majewska, B., Nowakowska, A., & Rosiak E. (2014). Evaluation of microbial quality of imported honeys available on the Warsaw market. In Proceedings of International Symposium on Bee Products 3rd Edition, Annual meeting of the International Honey Commission (IHC) (p. 117). Opatija – Croatia.

Midura, T.F., Snowden, S., Wood, R.M., Arnon SS. (1979). Isolation of Clostridium botulinum from honey. Journal of Clinical Microbiology, 9(2), 282-283.

Myllykoski, J., Nevas, M., Lindström, M., Korkeala, H. (2006). The detection and prevalence of Clostridium botulinum in pig intestinal samples. International Journal of Food Microbiology, 110(2),172-177. http://doi.org/10.1016/j.ijfoodmicro.2006.04.017

Nakano, H., Kizaki, H., & Sakaguchi G. (1994). Multiplication of Clostridium botulinum in dead honey-bees and bee pupae, a likely source of heavy contamination of honey. International Journal of Food Microbiology, 21(3), 247-252. http://doi.org/10.1016/0168-1605(94)90031-0

Nevas, M., Lindström, M., Hautamäki, K., Puoskari, S., Korkeala, H. (2005). Prevalence and diversity of Clostridium botulinum types A, B, E and F in honey produced in the Nordic countries. International Journal of Food Microbiology, 105(2), 145-151. http://doi.org/10.1016/j.ijfoodmicro.2005.04.007

Nevas, M. (2006). Clostrıdium botulinum in honey production with respect to infant botulism. Academic Dissertation, Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, University of Helsinki, Helsinki, Finland, Retrieved May 1, 2016, from https://helda.helsinki.fi/bitstream/handle/10138/18964/clostrid.pdf

Nevas, M., Lindström, M., Hörman, A., Keto-Timonen, R., Korkeala H. (2006). Contamination routes of Clostridium botulinum in the honey production environment. Environmental Microbiology, 8(6), 1085-1094. http://doi.org/10.1111/j.1462-2920.2006.01000.x

Palacio, A.M., Figini, E.F., Ruffinengo, S.R., Rodriguez, E.M., del Hoyo, M.L., Bedascarrasbure, E.L. (2000). Changes in a population of Apis mellifera L. selected for hygienic behaviour and its relation to brood disease tolerance. Apidologie, 31(4), 471-478. http://doi.org/10.1051/apido:2000139

PN-88/A-77626. (1988). Miód pszczeli. Warszawa: Wydawnictwo Normalizacyjne Alfa.

Roman, A., Popiela-Pleban, E., & Kozak M. (2013). Factors influencing consumer behavior relating to the purchasing of honey. Part 1. The buying process and the level of consumption. Journal of Apicultural Science, 57(2), 159-172. http://doi.org/10.2478/JAS-2013-0026

Różańska. H. (2011). Microbiological quality of Polish honey. Bulletin of the Veterinary Institute in Pulawy, 55(3), 443-445.

Rudnicka, K., Tenderenda, M., & Chmiela, M, (2013). Etiologia i epidemiologia botulizmu niemowląt. Pediatria Polska, 89(3),198-202. http://doi:10.1016/j.pepo.2013.10.003

Rudnicka, K., Kwiatkowska, P., Gajewski A., Chmiela M., (2015). Mikroflora miodu jako źródło spor C. botulinum i przyczyna rozwoju botulizmu niemowląt – rozważania na temat zasadności oczyszczania miodu w kontekście obowiązującego prawa. Postępy Mikrobiologii, 54(2), 184-194.

Satorres, S.E., Alcaraz, L.E., Fernández, R.A., Centorbi, O.N. (1999). Isolation of Clostridium botulinum in medical plants. Anaerobe, 5(3), 173-175. http://doi.org/10.1006/anae.1999.0205

Shapiro, R.L., Hatheway C., & Swerdlow D.L. (1998). Botulism in the United States. A clinical and epidemiologic review. Annals of Internal Medicine, 129(3), 221-228. http://doi.org/10.7326/0003-4819-129-3-199808010-00011

Sharma, S. K., Ferreira, J.L., Eblen, B.S., Whiting R.C. (2006). Detection of type A, B, E, and F Clostridium botulinum neurotoxins in foods by using an amplified enzyme-linked immunosorbent assay with digoxigenin-labeled antibodies. Applied Environmental Microbiology, 72(2), 1231–1238. http://doi.org/10.1128/AEM.72.2.1231-1238.2006

Smith, LD. (1979). Clostridium botulinum: characteristics and occurrence. Clinical Infectious Diseases, 1(4), 637-641. http://doi.org/10.1093/clinids/1.4.637

Snowdon, J. A., & Cliver, D.O. (1996). Microorganisms in honey. International Journal of Food Microbiology, 31(1-3), 1-26. http://doi.org/10.1016/0168-1605(96)00970-1

Spivak, M., & Downey, D.L. (1998). Field assays for hygienic behavior in honey bees (Hymenoptera: Apidae). Journal of Economic Entomology, 91(1), 64-70. http://doi.org/10.1093/jee/91.1.64.

Stämpfli, H.R. (2014). Botulism. In The Merck Veterinary Manual. Retrieved May 1, 2016, from http://www.merckvetmanual.com/mvm/index.html

Volkova, V.P., & Siniak, K.M. (1989). The possibility of the germination of spores of pathogenic clostridia and bacilli in soil. Izvestiia Akademii Nauk SSSR. Seriia Biologicheskaia, 5, 791-795.

Wells, C.L., & Wilkins, T.D. (1996). Clostridia: Sporeforming Anaerobic Bacilli. In Medical Microbiology. Texas: Galveston, Retrieved May 1, 2016, from http://www.ncbi.nlm.nih.gov/books/NBK8219/

Woudstra, C., Skarin, H., Anniballi, F., Fenicia, L., Bano, L., Drigo, I., Koene, M., Bäyon-Auboyer, M.H., Buffereau, J.P., De Medici, D., Fach P. (2012). Neurotoxin gene profiling of Clostridium botulinum types C and D native to different countries within Europe. Applied and Environmental Biology, 78(9), 3120–3127. http://doi.org/10.1128/AEM.07568-11

WHO (2013). Botulism. Fact sheet No 270. Retrieved May 1, 2016,, from http://www.who.int/mediacentre/factsheets/fs270/en/

Yule, A.M., Barker, I.K., Austin, J.W., Moccia, R.D. (2006). Toxicity of Clostridium botulinum type E neurotoxin to Great Lakes fish: implications for avian botulism. Journal of Wildlife Diseases, 42(3), 479-93. http://dx.doi.org/10.7589/0090-3558-42.3.479

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