Lactic acid bacteria (LAB) can cause deterioration of food quality even at low temperatures. In this study, we investigated the cold-adaptation mechanism of a novel food spoilage LAB, Leuconostoc mesenteroides NH04 (NH04). L. mesenteroides was isolated from several spoiled cooked meat products at a high frequency in our factories. NH04 grew rapidly at low temperatures within the shelf-life period and resulted in heavy financial losses. NH04 grew more rapidly than related strains such as Leuconostoc mesenteroides NBRC3832 (NBRC3832) at 10°C. Proteome analysis of NH04 demonstrated that this strain produces a homolog of alkyl hydroperoxide reductase––AhpC––the expression of which can be induced at low temperatures. The expression level of AhpC in NH04 was approximately 6-fold higher than that in NBRC3832, which was grown under the same conditions. Although AhpC is known to have an anti-oxidative role in various bacteria by catalyzing the reduction of alkyl hydroperoxide and hydrogen peroxide, the involvement of AhpC in cold adaptation of food spoilage bacteria was unclear. We introduced an expression plasmid containing ahpC into NBRC3832, which grows slower than NH04 at 10°C, and found that expression of AhpC enhanced growth. These results demonstrated that AhpC, which likely increases anti-oxidative capacity of LAB, plays an important role in their rapid growth at low temperatures.
To simplify the labor-intensive conventional routine testing of samples to detect Leuconostoc at a meat processing plant, we developed polymerase chain reaction (PCR) primers specific for Leuconostoc from 16S rRNA gene sequences. These primers did not detect other common lactic acid bacteria such as Lactobacillus plantarum, Lact. sake, Lact. fermentum, Lact. acidophilus and Weissella viridescens. PCR with this primer detected all Leuconostoc species tested (Leu. mesenteroides subsp. mesenteroides, Leu. pseudomesenteroides, Leu. carnosum, Leu. lactic, Leu. citreum, Leu. amelibiosum, Leu. gelidum), except for Leu. fallax, and no other lactic acid bacteria on agarose gel electrophoresis. The method could identify areas contaminated with Leuconostoc in a large-scale industrial meat processing plant. Of 69 samples analyzed, 34 were positive for Leuconostoc according to the conventional culture method (isolation of LAB producing dextran) and PCR, whereas 29 were negative according to both. Six samples were culture-negative but positive by PCR. No false negative results were generated by PCR. The method is rapid and simple, is useful for routinely monitoring areas contaminated with Leuconostoc in meat processing plants, and could help to prevent the spoilage of meat products.
To clarify the effects of water activity (a W ) and sodium lactate on the growth of Listeria monocytogenes in raw ham, an unheated processed meat product, L. monocytogenes ATCC 49594 (serotype 4b, Scott A) was inoculated into raw ham and its growth was investigated in five laboratories. L. monocytogenes did not grow in raw ham with an a W of 0.93 (0.930≤a W <0.940) in any of the laboratories during storage for 56 days at 10℃. It is presumed that an a W of 0.93 in raw ham acts additively and/or synergistically with the pH of meat, salt content, nitrite and low-temperature storage (10℃) to inhibit the growth of L. monocytogenes. On the other hand, in raw ham with an a W of 0.94 (0.940≤a W <0.950), L. monocytogenes growth was completely inhibited by the addition of 2 % sodium lactate. As a result, utilization of sodium lactate in raw ham produced in accordance with the Japanese Food Sanitation Law is valuable for the inhibition of L. monocytogenes growth.
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