Challenge Testing the Lactoperoxidase System Against a Range of Bacteria Using Different Activation Agents

Abstract 

Lactoperoxidase (LP) exerts antimicrobial effects in combination with H₂O₂ and either thiocyanate (SCN⁻) or a halide (e.g., I⁻). Garlic extract in the presence of ethanol has also been used to activate the LP system. This study aimed to determine the effects of 3 LP activation systems (LP+SCN⁻+H₂O₂; LP+I⁻+H₂O₂; LP + garlic extract + ethanol) on the growth and activity of 3 test organisms (Staphylococcus aureus, Pseudomonas aeru-ginosa, and Bacillus cereus). Sterilized milk was used as the reaction medium, and the growth pattern of the organisms and a range of keeping quality (KQ) indicators (pH, titratable acidity, ethanol stability, clot on boiling) were monitored during storage at the respective optimum growth temperature for each organism. The LP+I⁻+ H₂O₂ system reduced bacterial counts below the detection limit shortly after treatment for all 3 organisms, and no bacteria could be detected for the duration of the experiment (35 to 55h). The KQ data confirmed that the milk remained unspoiled at the end of the experiments. The LP + garlic extract + ethanol system, on the other hand, had no effect on the growth or KQ with P. aeruginosa, but showed a small retardation of growth of the other 2 organisms, accompanied by small increases (5 to 10h) in KQ. The effects of the LP+SCN⁻+H₂O₂ system were intermediate between those of the other 2 systems and differed between organisms. With P. aeruginosa, the system exerted total inhibition within 10h of incubation, but the bacteria regained viability after a further 5h, following a logarithmic growth curve. This was reflected in the KQ indicators, which implied an extension of 15h. With the other 2 bacterial species, LP+SCN⁻+H₂O₂ exerted an obvious inhibitory effect, giving a lag phase in the growth curve of 5 to 10h and KQ extension of 10 to 15h. When used in combination, I⁻ and SCN⁻ displayed negative synergy.

Key words: challenge testing, lactoperoxidase, antimicrobial effect, keeping quality