Abstract
Potential pathogens of foodborne toxic infections – bacterial contaminants Bacillus cereus isolated from plant raw materials and food products from the Ukrainian region were investigated. When determining of the proportion of isolated bacilli from the plant samples, it was established that the epidemiologically significant microorganisms of Bacillus cereus as agents of food poisoning are the second largest. The average value of contaminated samples of Ukrainian plant raw materials and processed products with Bacillus cereus is 36,2 %. The ability of Bacillus cereus strains identified by a complex of morphological, tinctorial, cultural and biochemical properties, to produce specific emetic and enterotoxins was studied. Molecular genetic diagnosis and detection of the toxin-producing ability of isolated 42 Bacillus cereus strains showed both the possibility of their rapid identification and the presence of specific toxicity genes. Multiplex polymerase chain reaction (PCR) was carried out with specific primers to detect toxicity determined of various bacilli genes: nheA, hblD, cytK, cesВ. The distribution of toxigenic genes is significantly different among the Bacillus cereus isolates from various sources. The nheA, hblD and cytK enterotoxin genes were detected in 100, 83,3 and 61,9 % of the investigated strains of Bacillus cereus, respectively. The cesB gene encoding emetic toxin was detected in 4,8 % of strains. Molecular-genetic PCR-method confirmed that all the isolated strains belong to the Bacillus cereus group, and the ability to produce toxins can be attributed to five groups. The main toxins that produce the investigated Bacillus cereus strains were nhe and hbl enterotoxins encoded by the corresponding genes of nheA and hblD. The enterotoxic type of Bacillus cereus was predominant in Ukrainian region. Studies of domestic plant food raw materials and products have confirmed the need to improve microbiological control of product safety by introducing accelerated specific diagnostics of contaminants by molecular genetics methods.
Highlights
The results suggest that the examined dried products, fruit and berries were free of the emetic toxin but not free of enterotoxins and the distribution of enterotoxic genes is significantly different among the B
For the use of DNA of Paenibacillus polymyxa and P. macerans no amplification product was obtained. These finding revealed that nhe and hbl enterotoxins encoded by nheA and hblD genes were the major toxins among B. cereus investigated in this study and enterotoxic type of B. cereus was predominant in Ukrainian region
By determining of the proportion of isolated bacilli from the plant samples, it was found that microorganisms of B. cereus group as agents of food poisoning are the second largest and consist 10 – 31 % from their total bacilli count
Summary
Досліджено потенційні збудники харчових токсикоінфекцій – токсигенні бацилярні контамінанти Bacillus cereus, виділені з рослинної сировини і продукції харчової промисловості українського регіону. Середнє значення контамінованості Bacillus cereus зразків української рослинної сировини і продуктів її переробки становить 36,2 %. Вивчено здатність штамів Bacillus cereus, ідентифікованих за комплексом морфологічних, тінкторіальних, культуральних та біохімічних властивостей, продукувати характерні еметичний (блювотний) і ентеротоксини. Молекулярно-генетична діагностика і виявлення токсинпродукучої здатності виділених 42 штамів Bacillus cereus показали як можливість їх швидкої ідентифікації, так і наявність характерних генів токсичності. Молекулярно-генетичним ПЛР-методом підтверджено, що всі виділені штами відносяться до групи Bacillus cereus, а за здатністю виробляти токсини їх можна віднести до п'яти груп. Які продукують досліджені штами Bacillus cereus, є ентеротоксини nhe та hbl, кодовані відповідними генами nheA та hblD. Ключові слова: токсинпродукуючі Bacillus cereus, ентеротоксини, еметичний токсин, молекулярно-генетична діагностика, полімеразна ланцюгова реакція, безпека харчових продуктів
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