The food industry seeks alternatives to satisfy consumer demands of safe foods with a long shelf-life able to maintain the nutritional and organoleptic quality. The application of antimicrobial compounds-producing protective cultures may provide an additional parameter of processing in order to improve the safety and ensure food quality, keeping or enhancing its sensorial characteristics. In addition, strong evidences suggest that certain probiotic strains can confer resistance against infection with enteric pathogens. Several mechanisms have been proposed to support this phenomenon, including antimicrobial compounds secreted by the probiotics, competitive exclusion, or stimulation of the immune system. Recent research has increasingly demonstrated the role of antimicrobial compounds as protective mechanism against intestinal pathogens and therefore certain strains could have an effect on both the food and the gut. In this aspect, the effects of the combination of different strains keep unknown. The development of multistrain probiotic dairy products with good technological properties and with improved characteristics to those shown by the individual strains, able to act not only as protective cultures in foods, but also as probiotics able to exert a protective action against infections, has gained increased interest.
The inhibitory activity of nisin (N), reuterin (R), and the lactoperoxidase system (LPS), added individually or in combination, against Listeria monocytogenes and Staphylococcus aureus was investigated in "cuajada" (curdled milk), a semisolid dairy product manufactured in Spain. Cuajada was manufactured from UHT skim milk separately inoculated with L. monocytogenes and Staph. aureus, each at approximately 4 log cfu/mL, and held under conditions of temperature abuse (10 degrees C). On d 3, a synergistic bactericidal activity was observed for the combinations of biopreservatives assayed, with L. monocytogenes counts of only 0.30 log cfu/mL in cuajada made with N + R + LPS vs. 8.31 log cfu/mL in control cuajada. After 12 d, L. monocytogenes could not be detected in cuajada made with added N + LPS or N + R + LPS. Staphylococcus aureus was more resistant than L. monocytogenes to biopreservatives added individually. On d 3, the synergistic effect of the 3 biopreservatives against Staph. aureus resulted in counts of 3.03 log cfu/mL in cuajada made with N + R + LPS vs. 6.40 in control cuajada. After 12 d, Staph. aureus counts were 2.61 log cfu/mL in cuajada made with N + R + LPS, whereas they ranged from 6.11 to 7.70 log cfu/mL in control cuajada and in cuajada made with other combinations of biopreservatives. The most pronounced decrease in pathogen counts was achieved by the triple combination N + R + LPS, which acted synergistically on the inactivation of L. monocytogenes and Staph. aureus in cuajada over 12 d at 10 degrees C. The treatment combining these 3 natural biopreservatives at low concentrations, within the hurdle concept of food preservation, might be a useful tool to control the growth of pathogenic microorganisms in nonacidified dairy products.
The ability of Listeria monocytogenes to grow at low temperatures and increase to
hazardous levels during storage of dairy products is of great concern to the dairy
industry. List. monocytogenes was detected in 3·6% of 774 cows' milk samples from
114 farms in Central Spain (Gaya et al. 1998). Ewes' and goats' milks produced in
Spain had contamination levels of 2·2 and 2·6% respectively (Rodríguez et al. 1994;
Gaya et al. 1996). List. monocytogenes may survive during the manufacture and
ripening of most cheese varieties, with higher risk in soft surface-ripened cheeses
owing to the rise in pH during ripening (Maisnier-Patin et al. 1992; Giraffa &
Carminati, 1997).The efficacy with which actively growing bacteriocin-producing cultures inhibit
List. monocytogenes in some cheese varieties has been demonstrated. Nisin-producing
starters inhibited List. monocytogenes V7 in Camembert cheese (Maisnier-Patin et al.
1992) and List. innocua BL86/26 in raw ewes' milk Manchego cheese (Rodríguez et al.
1998). A lacticin 3147-producing starter culture (McAuliffe et al. 1999) reduced
List. monocytogenes Scott A levels in cottage cheese. Inoculation of milk with
enterocin AS-48-producing Enterococcus faecalis INIA 4 inhibited List. monocytogenes
strains Ohio and Scott A during the manufacture and ripening of Manchego cheese
(Nuñez et al. 1997).Semi-hard cheese varieties from raw or pasteurized cows' milk mixed with
different proportions of ewes' and/or goats' milk represent ∼ 50% of Spanish cheese
production. The objective of the present work was to evaluate the antilisterial
activity of three bacteriocin-producing lactic acid bacteria strains isolated from milk
(Rodríguez et al. 2000) during the manufacture and ripening of a semi-hard cheese
manufactured from raw milk.
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