Bacteriocins and novel bacterial strains.

Authors: Stern, Norman; SVETOCH, E - State Research Center For Applied Microbiology And Biotechnology; ERUSLANOV, B - State Research Center For Applied Microbiology And Biotechnology; PERELYGIN, V - State Research Center For Applied Microbiology And Biotechnology; LEVCHUK, V - State Research Center For Applied Microbiology And Biotechnology; URAKOVQ, N - State Research Center For Applied Microbiology And Biotechnology; VOLODINA, L - State Research Center For Applied Microbiology And Biotechnology; KOVALEV, Y - State Research Center For Applied Microbiology And Biotechnology; KUDRYAVTESAVA, T - State Research Center For Applied Microbiology And Biotechnology; POKHILENKO, V - State Research Center For Applied Microbiology And Biotechnology; BORZENKOV, V - State Research Center For Applied Microbiology And Biotechnology; SVETOCH, O - State Research Center For Applied Microbiology And Biotechnology; MITSEVICH, E - State Research Center For Applied Microbiology And Biotechnology; MITSEVICH, I - State Research Center For Applied Microbiology And Biotechnology

Submitted to: Journal of Food Protection
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/15/2010
Publication Date: 6/22/2010
Citation: Stern, N.J., Svetoch, E.A., Eruslanov, B.V., Perelygin, V.V., Levchuk, V.P., Urakovq, N., Volodina, L.I., Kovalev, Y.N., Kudryavtesava, T.V., Pokhilenko, V.D., Borzenkov, V.N., Svetoch, O.E., Mitsevich, E.V., Mitsevich, I.P. 2010. Bacteriocins and novel bacterial strains. Journal of Food Protection. Vol 68,p11-17.

Interpretive Summary: Campylobacter causes a tremendous number of human diseases. The disease is thought to be transmitted through poultry. We sought and found an approach to control the large numbers of Campylobacter presently found on processed broilers. We identified several Bacillus bacteria that were antagonistic to Campylobacter. We then isolated the protein substances (bacteriocins) that were responsible for the antagonism. We characterized each of the substances and sequenced the amino acid structure to discriminate these from all other antimicrobial proteins. Using these bacteriocins we reduced colonized Campylobacter in chickens by one-million fold. Using these bacteriocins as therapies now provides resolution to important health problems associated with poultry.

Technical Abstract: Poultry is thought to be a significant source of Campylobacter in human disease. We evaluated anti-Campylobacter activity among 365 Bacillus and Paenibacillus isolates from poultry. One novel antagonistic Bacillus circulans and three Paenibacillus polymyxa strains were identified and further studied. Crude antimicrobial preparations were obtained. Zones of Campylobacter growth inhibition surrounding these preparations were quantified. These peptides were purified using combinations of molecular mass resolution and ion exchange chromatography. Molecular masses of the peptides were estimated at approximately 3,500 Da by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Isoelectric focusing was used to determine the pI values of the peptides. Amino acid sequences of the bacteriocins and more precise molecular masses were obtained by MALDI-TOF analysis. The bacteriocin from P. polymyxa NRRL B-30507 had a pI of 4.8, that from P. polymyxa NRRL B-30509 had a pI of 7.2, that from P. polymyxa NRRL B-30508 had a pI of 4.8, and that from B. circulans NRRL B-30644 had a pI of 7.8. The amino acid sequences were consistent with those of class IIa bacteriocins. These antagonists and the corresponding bacteriocins were useful in the control of Campylobacter infection in poultry. We evaluated a bacteriocin-based treatment to reduce Campylobacter jejuni colonization in poultry. In each of the eight trials, significant reductions in colonization by C. jejuni were observed (P < 0.05). Bacteriocin treatment dramatically reduced both intestinal levels and frequency of chicken colonization by C. jejuni.