Bio-protective potential of lactic acid bacteria: Effect of Lactobacillus sakei and Lactobacillus curvatus on changes of the microbial community in vacuum-packaged chilled beef원문보기
Objective: This study was to determine the bacterial diversity and monitor the community dynamic changes during storage of vacuum-packaged sliced raw beef as affected by Lactobacillus sakei and Lactobacillus curvatus. Methods: L. sakei and L. curvatus were separately incubated in vacuumed-packaged r...
Objective: This study was to determine the bacterial diversity and monitor the community dynamic changes during storage of vacuum-packaged sliced raw beef as affected by Lactobacillus sakei and Lactobacillus curvatus. Methods: L. sakei and L. curvatus were separately incubated in vacuumed-packaged raw beef as bio-protective cultures to inhibit the naturally contaminating microbial load. Dynamic changes of the microbial diversity of inoculated or non-inoculated (control) samples were monitored at $4^{\circ}C$ for 0 to 38 days, using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Results: The DGGE profiles of DNA directly extracted from non-inoculated control samples highlighted the order of appearance of spoilage bacteria during storage, showing that Enterbacteriaceae and Pseudomonas fragi emerged early, then Brochothrix thermosphacta shared the dominant position, and finally, Pseudomonas putida showed up became predominant. Compared with control, the inoculation of either L. sakei or L. curvatus significantly lowered the complexity of microbial diversity and inhibited the growth of spoilage bacteria (p<0.05). Interestingly, we also found that the dominant position of L. curvatus was replaced by indigenous L. sakei after 13 d for L. curvatus-inoculated samples. Plate counts on selective agars further showed that inoculation with L. sakei or L. curvatus obviously reduced the viable counts of Enterbacteraceae, Pseudomonas spp. and B. thermosphacta during later storage (p<0.05), with L. sakei exerting greater inhibitory effect. Inoculation with both bio-protective cultures also significantly decreased the total volatile basic nitrogen values of stored samples (p<0.05). Conclusion: Taken together, the results proved the benefits of inoculation with lactic acid bacteria especially L. sakei as a potential way to inhibit growth of spoilage-related bacteria and improve the shelf life of vacuum-packaged raw beef.
Objective: This study was to determine the bacterial diversity and monitor the community dynamic changes during storage of vacuum-packaged sliced raw beef as affected by Lactobacillus sakei and Lactobacillus curvatus. Methods: L. sakei and L. curvatus were separately incubated in vacuumed-packaged raw beef as bio-protective cultures to inhibit the naturally contaminating microbial load. Dynamic changes of the microbial diversity of inoculated or non-inoculated (control) samples were monitored at $4^{\circ}C$ for 0 to 38 days, using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Results: The DGGE profiles of DNA directly extracted from non-inoculated control samples highlighted the order of appearance of spoilage bacteria during storage, showing that Enterbacteriaceae and Pseudomonas fragi emerged early, then Brochothrix thermosphacta shared the dominant position, and finally, Pseudomonas putida showed up became predominant. Compared with control, the inoculation of either L. sakei or L. curvatus significantly lowered the complexity of microbial diversity and inhibited the growth of spoilage bacteria (p<0.05). Interestingly, we also found that the dominant position of L. curvatus was replaced by indigenous L. sakei after 13 d for L. curvatus-inoculated samples. Plate counts on selective agars further showed that inoculation with L. sakei or L. curvatus obviously reduced the viable counts of Enterbacteraceae, Pseudomonas spp. and B. thermosphacta during later storage (p<0.05), with L. sakei exerting greater inhibitory effect. Inoculation with both bio-protective cultures also significantly decreased the total volatile basic nitrogen values of stored samples (p<0.05). Conclusion: Taken together, the results proved the benefits of inoculation with lactic acid bacteria especially L. sakei as a potential way to inhibit growth of spoilage-related bacteria and improve the shelf life of vacuum-packaged raw beef.
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문제 정의
sakei as shown in the DGGE profile plus other LAB. These findings provided basic information for the further study about the interactions between inoculated and indigenous LAB cultures, and assist in the selection of better bio-protective cultures for raw meat.
제안 방법
The vacuum bags were composed of polyamide/ polyethylene film with oxygen permeability <15 cm3 /m2 /24 h at 25℃, 1 atm, and thickness of 80 μm. All the samples were collected immediately after inoculation (day 0) and at 7, 13, 21, 28, 38 days storage at 4℃. Three packages of Ls, Lc, or control cuts were used for the following analyses at each time interval.
curvatus, using the polymerase chain reaction-denaturing gradient gel electrophoresis technique (PCR-DGGE), and to make a comparison between the two strains to select a better bio-culture for raw beef. Moreover, the dominant bacteria at each storage time will be determined by conventional plate count methods, compared to analysis of the main bands on DGGE profiles, to verify the consistency of culture-dependent and independent methods. Also the pH value, color and total volatile basic nitrogen (TVBN) was measured to examine beef quality and stability upon addition of bio-preservatives.
The effects of L. sakei and L. curvatus on the microbial load (TVC, total LAB, Enterbacteraceae, Pseudomonas pp. and B. thermosphacta) and beef quality during extended storage were evaluated using the analysis of variance using IBM SSPS Statistic (19.0.0). All the data were represented as mean±standard deviation.
Also, in spite of a large literature on the subject, there are few studies focusing on the comparable antibacterial properties of different LAB strains on raw beef, which is very important to select the most suitable strains for a specific food. Thus, the aim of this study was to determine the bacterial diversity and monitor the community dynamic changes during storage of vacuum-packaged sliced raw beef as affected by L. sakei or L. curvatus, using the polymerase chain reaction-denaturing gradient gel electrophoresis technique (PCR-DGGE), and to make a comparison between the two strains to select a better bio-culture for raw beef. Moreover, the dominant bacteria at each storage time will be determined by conventional plate count methods, compared to analysis of the main bands on DGGE profiles, to verify the consistency of culture-dependent and independent methods.
대상 데이터
Sample procedures: The beef cattle slaughter procedure was performed according to the guidelines of the Animal Ethics Committee in Shandong Agricultural University as approved by the State Scientific and Technological Commission in November 1988 (China). Twelve conventionally segmented longissimus lumborum (LL, 12th rib to the last lumbar vertebrae, 24 h postmortem, pH24h 5.4 to 5.7) from six carcasses were selected from a cattle slaughter plant based in Shandong province, China. Muscles from the same animal were pairvacuum packed, placed in a foam box and transported to the laboratory within 2 h.
이론/모형
The filtrates were refrigerated until further use. The TVBN content was measured by semi-micro diffusion method [24,25].
성능/효과
Additionally, a distinctive result we found was that the dominant position of L. curvatus was replaced by indigenous L. sakei after 13 days for L. curvatus-inoculated samples (Figure 1). Castellano et al [30] reported that the bio-protective culture L.
Microbial diversity of vacuum-packaged raw beef during the 38 days storage at 4°C (control samples): There was a high initial bacterial diversity in the vacuum-packaged raw beef. Seven bands were detected at 0 d in the control samples, of which band 2 was the most intensive one, suggesting that the corresponding bacteria for this band was the main bacterial contaminant from the slaughter plant, and it was defined as L. sakei according to the sequencing results (Table 1). Bands 1, 3, and 9 in lane C0 were some uncultured bacteria and uncommon contaminating bacteria in raw beef, while bands 12 and 13 were very common bacteria found in vacuum-packaged meat, belonging to LAB.
The study of Katikou et al [6] also found LAB treated beef samples were not significantly different with untreated beef, and the L* value showed an increasing trend. Taken together, results indicated that the addition of L. sakei and L. curvatus bio-protective cultures on raw beef samples had no negative effect on the beef color, and on the contrary, it seems that L. sakei could improve the L* value in the latter stage of storage.
curvatus) as bio-protective cultures in vacuumed-packaged raw beef at 4℃ were evaluated by using both culture-independent and culture-dependent methods. The PCR-DGGE results demonstrated that the inoculation of either L. sakei or L. curvatus significantly lowered the complexity of microbial diversity and slowed down the growth of spoilage bacteria during the whole storage time, which coincided with a marked reduction of the viable counts of Enterbacteraceae, Pseudomonas spp. and B.
The final pH values of samples incubated with L. sakei and L. curvatus were 5.27 and 5.34, respectively, significantly (p<0.05) lower than the control sample (5.66).
Uncultured Lactobacillus, Lactobacillus fuchuensis, and Leuconostoc carnosum corresponded to bands 14, 15, and 16, respectively, together with Lactobacillus graminis and Leuconostoc mesenteroides (bands 12 and 13), were not significantly influenced by the bio-protective cultures, as those bands were only detected at trace levels in most of the samples across the entire storage period.
후속연구
sakei inoculation treatment. Future efforts should also be oriented towards expanding our knowledge regarding the interaction between inoculated bio-protective cultures and the indigenous LAB in raw beef.
참고문헌 (38)
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