Isolation of Bacillus circulans and Paenibacillus polymyxa Strains Inhibitory to Campylobacter jejuni and Characterization of Associated Bacteriocins

Abstract

We evaluated anti-Campylobacter activity among 365 Bacillus and Paenibacillus isolates from poultry production environments. One novel antagonistic Bacillus circulans and three Paenibacillus polymyxa strains were identified and further studied. Cell-free ammonium sulfate precipitate (crude antimicrobial preparation) was obtained from each candidate culture. Zones of Campylobacter growth inhibition surrounding 10 microl of this crude antimicrobial preparation were quantified using a spot test. Campylobacter growth resumed when the preparation was preincubated with selected protease enzymes, demonstrating peptide characteristics consistent with a bacteriocin. These peptides were further 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 matrix-assisted laser desorption and ionization-time of flight (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 may be useful in the control of Campylobacter infection in poultry.

In Vitro Antagonistic Activity Evaluation of Lactic Acid Bacteria (LAB) Combined with Cellulase Enzyme Against Campylobacter jejuni Growth in Co-Culture

“…In our experimental conditions, Paenibacillus sp. CWBI B1070 did not exert any anti-Campylobacter activity, in contrast to the results of Svetoch et al [33], who reported antagonistic activities against Campylobacter of one Bacillus and three Paenibacillus strains isolated from the poultry production environment.…”

Production of a newly isolated Paenibacillus polymyxa biocontrol agent using monosodium glutamate wastewater and potato wastewater

“…polymyxa has also been isolated from several places such as food, rhizosphere, poultry production environments, and soils; and most of them can restrain fungal pathogen growth (He et al, 2007;Raza et al, 2009;Svetoch et al, 2005). Further studies will investigate the anti-fungal pathogen activity of the isolate EBL-06 both in phyllosphere and in rhizosphere by field trials.…”

Utilization of Sweat Potato Starch Wastewater and Monosodium Glutamate Wastewater for Cultivation of an Anti-Fungal Biocontrol Agent Paenibacillus Polymyxa

“…Compared with other culture media, the mixture of SPW and MGW is the most suitable for the biocontrol agent production from the economic and environmental point of view. P. polymyxa has also been isolated from several places such as food, rhizosphere, poultry production environments, soils and most of them can restrain fungal pathogen growth (He et al, 2007;Raza et al, 2009;Svetoch et al, 2005). Further studies will investigate the antifungal pathogen activity of the isolate EBL06 both in phyllosphere and in rhizosphere by field trials.…”