238 BOTULINUM NEUROTOXIN SEROTYPES INVOLVED IN FOOD-BORNE BOTULISM OUTBREAKS IN ROMANIA IN THE LAST FIVE YEARS Laurenţiu Mihai CIUPESCU 1 , Isabela Madălina NICORESCU 1 , Rodica DUMITRACHE 1 , Rodica TANASUICA 1 , Veronica CIUPESCU 2 1 Institute of Hygiene and Veterinary Public Health, 5 Campul Mosilor Street, District 2, 021201, Bucharest, Romania, Phone: +4021.252.46.51, Fax: + 4021.252.00.61, Email: ciupescu_laurentiu@yahoo.com, isabela_nicorescu@yahoo.com, rdumitrache@yahoo.com, rodica.tanasuica@iispv.ro 2 Sanitary Veterinary Directorate of Bucharest, Ilioara 16Y, District 3, 032135, Bucharest, Romania, Phone: +4021.348.05.65, Fax: + 4021.348.23.52, Email: ciupescu_veronica@yahoo.com Corresponding author email: ciupescu_laurentiu@yahoo.com Abstract This paper summarizes five food-borne botulism outbreaks occurred in Romania from January 2010 till the beginning of 2015. In this period, from 54 food samples received from human botulism suspicion cases and 140 self-control samples, only five samples were positive to the botulinum neurotoxin detection by mouse bioassay. Traditional prepared food seems to be the most common way to get the causative bacteria from specific poisoning areas. The food matrices positive for BoNT were pork and fish meat, all of them made at home in a traditional way. The most frequent BoNT serotype incriminated was B, found in three outbreaks associated with homemade salted and smoked dried pork and one outbreak with a homemade salted and smoked-dried fish meat. Only in one case, two BoNT serotypes A and B were detected in the same sample (salted and smoked- dried pork meat). For certain regions, seems to be incriminated a certain type of BoNT. Amongst the five outbreaks, three were reported in the North-Western, one in the Western and one in the Southern area of the country, thereby these places could be assigned like botulism poisoning zones, but studies need to be continued. Key words: botulism, outbreaks, BoNT serotypes, Romania. INTRODUCTION Botulinum neurotoxins (BoNTs) are produced by six anaerobic spore-forming Gram positive phylogenetically and physiologically distinct clostridia (Clostridium botulinum Groups I-IV and some strains of Clostridium baratii and Clostridium butyricum) (Peck, 2009). BoNTs are the most acutely lethal and powerful naturally occurring toxins known to science, leading to neuroparalytic illness by inhibition of acetylcholine release in synapses (Sharma and Whiting, 2005; Vossen et al., 2012). Food-borne botulism is rare but must be considered a life-threatening emergency, requiring rapid recognition of the disease and identification of the source and type of the toxin (CDC, 1998). Knowing the toxin type is important in selecting an antitoxin for treatment (antitoxin produced against one type is not protective against others). The causative bacteria and spores are ubiquitous in nature but the distribution of strains can vary with the geographic area. The bacteria/spores alone do not cause the disease, only their production of botulinum toxin in anaerobic conditions renders them pathogenic. In the last two decades, various BoNT detection methods appeared like ELISA (Abbasova et al., 2011), Endopep-MS (Hedeland et al., 2011), ELISA-PCR (Fach, 2002) but the “gold standard” is still the mouse bioassay (Quinn et al., 1994). There are seven types of BoNT recognized (A to G) and 32 subtypes (Barash and Arnon, 2014), but the prevalent in human botulism are A, B, E and F types (Barr et al., 2005). Most strains produce only one type of toxin, but strains producing multiple toxin types are not unprecedented (Gimenez and Gimenez, 1993; Santos-Buelga et al., 1998; Barash and Arnon, 2004). In the last few years, Romania has experienced several food-borne botulism outbreaks, as follows: two outbreaks with 27 human cases in Scientifc Works. Series C. Veterinary Medicine. Vol. LXI (2) ISSN 2065-1295; ISSN 2343-9394 (CD-ROM); ISSN 2067-3663 (Online); ISSN-L 2065-1295