真空包装烟熏火腿切片特定腐败菌及靶向抑制研究
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摘要
低温肉制品因其特有的营养成分和组织结构,如脂肪、蛋白质含量丰富、肉质鲜嫩,风味浓郁,很受消费者的青睐,成为未来肉制品发展的方向。但低温肉制品水分活度较高,pH值也相对较高,加之在加工过程中灭菌不彻底,在后期的储存和销售过程中,一旦条件适宜或受到污染,产品中残留的细菌就会迅速地生长繁殖,导致产品腐败,产生诸如褪色、出水、出油、发黏、风味差、胀袋等不同程度的质量问题。如何延长低温肉制品的货架期,是摆在许多肉制品生产企业面前的一道难题。因此,以低温肉制品腐败微生物控制技术为核心的品质控制技术和综合保鲜技术研究,成为低温肉制品研究与产业化的关键问题。本研究的目的是运用DNA指纹图谱技术等现代生物技术,结合传统的微生物分离技术,以典型低温肉制品—真空包装烟熏火腿切片为代表,分离、鉴别其中的主要腐败微生物菌相,揭示低温火腿切片中特定腐败菌(SSO)的动态消长规律,及其对产品品质的影响机制;对产品中腐败菌相的形成加以确切阐述,建立快速准确地确定低温肉制品中特定腐败菌的方法及主要污染源追溯方法,确定关键质量控制点(CCP)。并进一步研究应用生物保护菌靶向抑制低温肉制品中特定腐败菌的生物效应;最终,为低温肉制品的生产、贮藏和流通过程中的品质安全保障提供理论依据和技术支持。具体研究内容和结果如下:
1.真空包装烟熏火腿切片贮藏特性研究
对不同温度贮藏条件下真空包装烟熏火腿切片的贮藏特性进行研究。结果表明,真空包装烟熏火腿切片初始pH值为6.42,A_w值为0.982,蛋白含量16.8%,脂肪含量3.3%,水分含量70.3%,初始菌数为10~2～10~3CFU/g。产品很容易腐败变质。产品在贮藏过程中,pH值呈下降趋势,TVB-N值和可溶性蛋白呈上升趋势,L~*值下降,a~*、b~*值上升,A_w值、脂肪含量及TBARS值变化不明显。
贮藏温度对产品质量和货架期影响显著,低温可以延长产品货架期。产品腐败表现为:腥臭味,酸腐味,颜色淡泊无光泽,质地松散无弹性,出水,产黏和产气。腐败胀袋气体的主要成分是二氧化碳、此外还含有乙醇等醇类物质、乙酸等挥发性酸类化合物。
贮藏期间,微生物达到的最大菌落数量为10~8CFU/g,以此为指标判断产品的货架期与感官判断一致。乳酸菌是真空包装烟熏火腿切片中的主要微生物,其次是肠杆菌科菌、假单胞菌、葡萄球菌/微球菌占少数,产品中有酵母菌污染。
2.产品贮藏期间微生物种群动态变化及主要腐败菌菌相分析
采用DGGE技术和传统微生物培养相结合的方法,分析探讨真空包装烟熏火腿切片中污染细菌的多样性和贮藏过程中的动态变化并分离、鉴别其中主要的腐败菌菌相。结果表明,产品贮藏初期污染微生物具有很高的多样性,但贮藏过程中逐渐发展成为少数特定腐败菌生长占优势,它们具有很强的生长竞争力,抑制了其它微生物的生长,造成产品质量劣变的特征性腐败。
两种方法结合共分离出真空包装烟熏火腿切片中主要腐败菌,它们分别代表:清酒乳杆菌(Lactobacillus sakei),弯曲乳杆菌(Lactobacillus curvatus),明串珠菌属的肠膜明串珠菌(Leuconostoc mesenteroides)和非培养的明串珠菌(Uncultured Leuconostoc sp.)
3.腐败菌对真空包装烟熏火腿切片的质量影响
通过分析腐败细菌产生腐败现象的腐败潜能和其产生腐败代谢产物的活力,研究导致产品腐败的微生物效应。结果表明:
肠膜明串珠菌肠膜亚种(Leuconostoc mesenteroides subsp.mesenteroides)利用肉基质中的葡萄糖进行异型发酵产生二氧化碳、乙酸、丙醇、丁醇,并分解蛋白质等含氮化合物造成TVB-N值升高。是造成真空包装烟熏火腿切片“空皮”,出水、“胀袋”、结构松散、颜色灰白等腐败现象的特定腐败菌。
清酒乳杆菌清酒亚种(Lactobacillus sakei subsp.sakei)优先发酵肉制品中的糖类产生乳酸、乙酸等酸性物质,造成pH值下降;产生黏多糖(葡萄糖-半乳糖多聚糖),形成黏液;生成酪胺、腐胺等典型腐败物质,产生腐败味。是造成真空包装烟熏火腿切片发酸变味、发黏起丝等腐败现象的特定腐败菌。
弯曲乳杆菌蜜二糖亚种(Lactobacillus curvatus subsp.melibiosus)优先发酵肉制品中的糖类产生乳酸、乙酸、乙醇等物质,产生黏多糖(葡萄糖-半乳糖多聚糖),并产生大量尸胺、腐胺、酪胺等典型腐败物质。是造成真空包装烟熏火腿切片发酸变味、发黏起丝、颜色暗淡、腐败臭味的特定腐败菌。
4.加工过程中的微生物种群差异性分析
运用DGGE技术分析烟熏火腿切片加工过程中主要生产关键环节的微生物种群多样性,通过比较各个污染环节微生物种群的差异,得出它们与产品中主要腐败菌的相互关系,结果表明:真空包装烟熏火腿切片中主要腐败菌来源于原料肉。生产过程中容易发生污染的环节是滚揉和热处理后的冷却过程。
防腐剂使用不当会给产品带来安全隐患。
5.生物保护菌对特定腐败菌的靶向抑制作用研究
评价运用清酒乳杆菌B-2(Lactobacillus sakei B-2)作为生物保护菌对延长真空包装烟熏火腿切片在4℃贮存货架期的作用。通过描述微生物在肉制品低温贮藏期间的动态变化过程,阐述生物保护菌与抑制目标菌之间是否有相互作用。结果表明,真空包装烟熏火腿切片接种清酒乳杆菌B-2 5.91±0.04 log_(10) CFU/g,于4℃贮藏,货架期为35 d,对照样货架期为15 d。使用生物保护菌L.sakei B-2抑制了低温肉制品中的主要腐败菌清酒乳杆菌(L.sakei)弯曲乳杆菌(L.curvatus)和肠膜明串珠菌(Leuconostoc.mssenteroides),但是不能完全抑制非培养的明串珠菌(unculturedLeuconostoc)。
应用传统微生物分离培养方法和分子生物学方法清楚地阐明了生物保护菌对目标菌的抑制效果。PCR-DGGE技术作为一种有效的技术手段,可以直观、准确地分析生物保护菌应用于延长低温肉制品货架期,微生物种群的动态变化。更好地了解生物保护菌与目标菌之间的靶向抑制作用有利于生物保护菌在肉制品中的正确应用。
Pasteurized meat products are favor meat products of customers with its proper nutrition and texture such as: enriched fat and protein, fresh and tender of tast, great flavor et al. and will be the way of future development of meat products. But pasteurized meat product has higher A_w and pH value. In addition, pasteurization could not kill all bacteria of product in the processing. So the survival bacteria would growth rapidly when the condition is suitable for its development or have recontamination which will cause for spoilage and resulting in short shelf-life with sensory quality defects such as discolouration, water liquid, oil exudation, ropy slime formation, sour off-flavour, gas production et al. How to prolong the shelf-life of pasteurized meat products is a difficult problem towards the manufactures. Therefore, the study of quality cuntrol and integration protective technology focusing on controlling the spoilage bacteria in these products became the key issue of research and industrialization of pasteurized meat products. The purpose of present study was isolation and identification the predominate spoilage bacteria in one of the representative pasteurized meat products—vacuum packed sliced smoked cooked ham, understanding the dynamic changes in specific spoilage organisms (SSO) of this product, and the effect mechanism of the SSO on the quality of meat products; explaining the formation of spoilage bacteria community, establishing methods to quickly and accurately identify the SSO and trace the main contaminated source; moreover, studying the bioprotective culture target inhibit towards the SSO in pasteurized meat products was carried out; Finally, we would like to provide a theory basis and technique for the ensuring of the quality and safety of pasteurized meat products in the period of prosessing, storage and distribution. The contents and results are as follows:
1. Study of the storage characteristic of vacuum packed sliced smoked cooked ham
The storage characteristic of vacuum packed sliced smoked cooked ham under different temperature was studied. The result showed that the vacuum packed sliced smoked cooked ham is a highly perishable meat product with initial pH value 6.42, A_w 0.982, protain content 16.8%, fat content 3.3%, water content 70.3%, and initial total bacteria count of 10~1～10~3 CFU/g. The pH value and L showed downtrend, TVB-N value、solubility protein and a~*、b~* value showed uptrend, and A_w, fat and TBARS value have no changed in significantly during the period of storage.
The storage temperature had significant effect on the quality and shelf-life of this product which storaged under low temperature would prolong the shelf-life. The sensory defects of spoilage products were sour off-flavour, discolouration, lackluster, relax texture, loosen elasticity, water liquid, ropy slime formation and gas production. The main elements of spoilage gas include CO_2, alcoholic compound of ethanol, volatility compound of acetic acidetal.
The maximum total bacteria count reached 10~8 CFU/g during the period of storage. Using the maximum bacteria count as level to exitmate the shelf-life of this product in accord with sensory evaluation method. The lactobacillus spp. (LAB) was the predominate microorganisms in vacuum packed sliced smoked cooked ham, Enterobacteriaceae taken second place, Pseudomonas spp., Staphylococci/Micrococcus were in the minority, and sometimes contaminated yeast in products.
2. Study on the microbial community dynamic during the period of storage and isolation and identification of the predominate spoilage bacteria
The microbial diversity and dynamic changes during the storage were explored through PCR-DGGE fingerprinting technique combination of conventional culture method. The result showed that the diversity of the original contaminated microorganism were high. However, the minority specifie spoilage bacteria eventually became the predomination microorganisms during the period of storage. They inhibited the growth of other microorganisms with strong competition ability caused specific spoilage which resulting in bad quality.
The main spoilage bacteria were isolated and identified from vacuum packed sliced smoked cooked ham by combination of culture dependent and culture independent methods. They were: Lactobacillus sakei, Lactobacillus curvatus, Leuconostoc mesenteroides and Uncultured Leuconostoc sp.
3. Spoilage microbial effect on the quality changes of vacuum packed sliced smoked cooked ham
The study of microbial effect on products spoilage was carried out by analysised the spoilage potential of microorganism of its can result in the phenolmenal of spoilage and the spoilage activeity of microorganism of its quantitative ability to produce spoilage metabolites. The results showed that:
Leuconostoc mesenteroides subsp. mesenteroides was indentified as a SSO that contributed to the quality defect such as "shell off", water liquid, "swell bag", relax texture and discolouration, lackluster. It can utilize the glucose of meat medium to heterofermentate to produce CO_2, acetic acid, propylanol, butyricanol, decompose compound of nitrogen such as protein to make the TVB-N value rise up.
Lactobacillus sakei subsp. sakei was identified as a SSO that contributed to quality defects such as sour off-flavor, ropy slime formation. It has preferential fermentation ability to utilize the sugar of meat products to produce lactic acid, acetic acid resulting in pH value drop, exopolysaccharide compound to formate ropy slime, produce representative spoilage compound of tyramine and putrescine resulting in rotten flavour.
Lactobacillus curvatus subsp. melibiosus was identified as a SSO that contributed to the quality defects such as sour off-flavor, ropy slime formation, discoloruration and grayness, rotten flavorur. It has preferential fermentation ability to utilize the glucide of meat products to produce lactic acid, acetic acid and ethanol, and exopolysaccaride compound, furthermore, it can make a great deal of representative spoilage compound of tyramine、cadaverine and putrescine when meat products become spoilage.
4. Analysis of the diversity of microbial community during processing of smoked cooked ham
The diversity of microbial community of the key processing point was analysed by DGGE technique. Through compared the diversity of microbial community between every susceptibly contamination point, the correlation of these microbial community and predominate spoilage bacteria was obtained. The results indicated that:
The main spoilage bacteria of vacuum packed sliced smoked cooked ham are come from the raw meat.
The processing of tumbling and the period of chilling after heat treatment are susceptible contamination points.
Improper using preservative will bring underlying trouble to the product safety.
5. Study of protective effect of bioprotective culture towards the SSO
The effect of Lactobacillus sakei B-2 as bioprotective culture to prolong the shelf-life of sliced vacuum packed smoked cooked ham storaged under 4℃was evaluated. The inhibitory interaction between bioprotective culture and target bacteria was expatiated by characterization of the dynamoic changes of microbial community of meat products during storage under refrigement temperature. The result showed that: The shelf-life of sliced, vacuum packed cooked ham inoculated with L. sakei B-2 at levels of 5.91±0.04 log_(10) CFU/g was 35 days, compared to the 15 days shelf-life of the control product. Treatment with the protective culture has been shown to suppress the growth of the predominant spoilage bacteria including L.sakei, L.curvatus and Le.mesenteroides but can not completely control the growth of strains of uncultured Leuconostoc.
The application of culture-dependent and in-dependent methods led to an understanding of the inhibitory interaction taking place between protective cultures and target strains. PCR-DGGE is an effective technique to analyze the dynamic changes of microorganism population when bioprotective culture applied in pasteurized meat products to prolong the shepf-life. A better understanding of inhibitory interaction between protective cultures and target strains may lead to correct application of protective cultures to meat products.
1.真空包装烟熏火腿切片贮藏特性研究
对不同温度贮藏条件下真空包装烟熏火腿切片的贮藏特性进行研究。结果表明,真空包装烟熏火腿切片初始pH值为6.42,A_w值为0.982,蛋白含量16.8%,脂肪含量3.3%,水分含量70.3%,初始菌数为10~2～10~3CFU/g。产品很容易腐败变质。产品在贮藏过程中,pH值呈下降趋势,TVB-N值和可溶性蛋白呈上升趋势,L~*值下降,a~*、b~*值上升,A_w值、脂肪含量及TBARS值变化不明显。
贮藏温度对产品质量和货架期影响显著,低温可以延长产品货架期。产品腐败表现为:腥臭味,酸腐味,颜色淡泊无光泽,质地松散无弹性,出水,产黏和产气。腐败胀袋气体的主要成分是二氧化碳、此外还含有乙醇等醇类物质、乙酸等挥发性酸类化合物。
贮藏期间,微生物达到的最大菌落数量为10~8CFU/g,以此为指标判断产品的货架期与感官判断一致。乳酸菌是真空包装烟熏火腿切片中的主要微生物,其次是肠杆菌科菌、假单胞菌、葡萄球菌/微球菌占少数,产品中有酵母菌污染。
2.产品贮藏期间微生物种群动态变化及主要腐败菌菌相分析
采用DGGE技术和传统微生物培养相结合的方法,分析探讨真空包装烟熏火腿切片中污染细菌的多样性和贮藏过程中的动态变化并分离、鉴别其中主要的腐败菌菌相。结果表明,产品贮藏初期污染微生物具有很高的多样性,但贮藏过程中逐渐发展成为少数特定腐败菌生长占优势,它们具有很强的生长竞争力,抑制了其它微生物的生长,造成产品质量劣变的特征性腐败。
两种方法结合共分离出真空包装烟熏火腿切片中主要腐败菌,它们分别代表:清酒乳杆菌(Lactobacillus sakei),弯曲乳杆菌(Lactobacillus curvatus),明串珠菌属的肠膜明串珠菌(Leuconostoc mesenteroides)和非培养的明串珠菌(Uncultured Leuconostoc sp.)
3.腐败菌对真空包装烟熏火腿切片的质量影响
通过分析腐败细菌产生腐败现象的腐败潜能和其产生腐败代谢产物的活力,研究导致产品腐败的微生物效应。结果表明:
肠膜明串珠菌肠膜亚种(Leuconostoc mesenteroides subsp.mesenteroides)利用肉基质中的葡萄糖进行异型发酵产生二氧化碳、乙酸、丙醇、丁醇,并分解蛋白质等含氮化合物造成TVB-N值升高。是造成真空包装烟熏火腿切片“空皮”,出水、“胀袋”、结构松散、颜色灰白等腐败现象的特定腐败菌。
清酒乳杆菌清酒亚种(Lactobacillus sakei subsp.sakei)优先发酵肉制品中的糖类产生乳酸、乙酸等酸性物质,造成pH值下降;产生黏多糖(葡萄糖-半乳糖多聚糖),形成黏液;生成酪胺、腐胺等典型腐败物质,产生腐败味。是造成真空包装烟熏火腿切片发酸变味、发黏起丝等腐败现象的特定腐败菌。
弯曲乳杆菌蜜二糖亚种(Lactobacillus curvatus subsp.melibiosus)优先发酵肉制品中的糖类产生乳酸、乙酸、乙醇等物质,产生黏多糖(葡萄糖-半乳糖多聚糖),并产生大量尸胺、腐胺、酪胺等典型腐败物质。是造成真空包装烟熏火腿切片发酸变味、发黏起丝、颜色暗淡、腐败臭味的特定腐败菌。
4.加工过程中的微生物种群差异性分析
运用DGGE技术分析烟熏火腿切片加工过程中主要生产关键环节的微生物种群多样性,通过比较各个污染环节微生物种群的差异,得出它们与产品中主要腐败菌的相互关系,结果表明:真空包装烟熏火腿切片中主要腐败菌来源于原料肉。生产过程中容易发生污染的环节是滚揉和热处理后的冷却过程。
防腐剂使用不当会给产品带来安全隐患。
5.生物保护菌对特定腐败菌的靶向抑制作用研究
评价运用清酒乳杆菌B-2(Lactobacillus sakei B-2)作为生物保护菌对延长真空包装烟熏火腿切片在4℃贮存货架期的作用。通过描述微生物在肉制品低温贮藏期间的动态变化过程,阐述生物保护菌与抑制目标菌之间是否有相互作用。结果表明,真空包装烟熏火腿切片接种清酒乳杆菌B-2 5.91±0.04 log_(10) CFU/g,于4℃贮藏,货架期为35 d,对照样货架期为15 d。使用生物保护菌L.sakei B-2抑制了低温肉制品中的主要腐败菌清酒乳杆菌(L.sakei)弯曲乳杆菌(L.curvatus)和肠膜明串珠菌(Leuconostoc.mssenteroides),但是不能完全抑制非培养的明串珠菌(unculturedLeuconostoc)。
应用传统微生物分离培养方法和分子生物学方法清楚地阐明了生物保护菌对目标菌的抑制效果。PCR-DGGE技术作为一种有效的技术手段,可以直观、准确地分析生物保护菌应用于延长低温肉制品货架期,微生物种群的动态变化。更好地了解生物保护菌与目标菌之间的靶向抑制作用有利于生物保护菌在肉制品中的正确应用。
Pasteurized meat products are favor meat products of customers with its proper nutrition and texture such as: enriched fat and protein, fresh and tender of tast, great flavor et al. and will be the way of future development of meat products. But pasteurized meat product has higher A_w and pH value. In addition, pasteurization could not kill all bacteria of product in the processing. So the survival bacteria would growth rapidly when the condition is suitable for its development or have recontamination which will cause for spoilage and resulting in short shelf-life with sensory quality defects such as discolouration, water liquid, oil exudation, ropy slime formation, sour off-flavour, gas production et al. How to prolong the shelf-life of pasteurized meat products is a difficult problem towards the manufactures. Therefore, the study of quality cuntrol and integration protective technology focusing on controlling the spoilage bacteria in these products became the key issue of research and industrialization of pasteurized meat products. The purpose of present study was isolation and identification the predominate spoilage bacteria in one of the representative pasteurized meat products—vacuum packed sliced smoked cooked ham, understanding the dynamic changes in specific spoilage organisms (SSO) of this product, and the effect mechanism of the SSO on the quality of meat products; explaining the formation of spoilage bacteria community, establishing methods to quickly and accurately identify the SSO and trace the main contaminated source; moreover, studying the bioprotective culture target inhibit towards the SSO in pasteurized meat products was carried out; Finally, we would like to provide a theory basis and technique for the ensuring of the quality and safety of pasteurized meat products in the period of prosessing, storage and distribution. The contents and results are as follows:
1. Study of the storage characteristic of vacuum packed sliced smoked cooked ham
The storage characteristic of vacuum packed sliced smoked cooked ham under different temperature was studied. The result showed that the vacuum packed sliced smoked cooked ham is a highly perishable meat product with initial pH value 6.42, A_w 0.982, protain content 16.8%, fat content 3.3%, water content 70.3%, and initial total bacteria count of 10~1～10~3 CFU/g. The pH value and L showed downtrend, TVB-N value、solubility protein and a~*、b~* value showed uptrend, and A_w, fat and TBARS value have no changed in significantly during the period of storage.
The storage temperature had significant effect on the quality and shelf-life of this product which storaged under low temperature would prolong the shelf-life. The sensory defects of spoilage products were sour off-flavour, discolouration, lackluster, relax texture, loosen elasticity, water liquid, ropy slime formation and gas production. The main elements of spoilage gas include CO_2, alcoholic compound of ethanol, volatility compound of acetic acidetal.
The maximum total bacteria count reached 10~8 CFU/g during the period of storage. Using the maximum bacteria count as level to exitmate the shelf-life of this product in accord with sensory evaluation method. The lactobacillus spp. (LAB) was the predominate microorganisms in vacuum packed sliced smoked cooked ham, Enterobacteriaceae taken second place, Pseudomonas spp., Staphylococci/Micrococcus were in the minority, and sometimes contaminated yeast in products.
2. Study on the microbial community dynamic during the period of storage and isolation and identification of the predominate spoilage bacteria
The microbial diversity and dynamic changes during the storage were explored through PCR-DGGE fingerprinting technique combination of conventional culture method. The result showed that the diversity of the original contaminated microorganism were high. However, the minority specifie spoilage bacteria eventually became the predomination microorganisms during the period of storage. They inhibited the growth of other microorganisms with strong competition ability caused specific spoilage which resulting in bad quality.
The main spoilage bacteria were isolated and identified from vacuum packed sliced smoked cooked ham by combination of culture dependent and culture independent methods. They were: Lactobacillus sakei, Lactobacillus curvatus, Leuconostoc mesenteroides and Uncultured Leuconostoc sp.
3. Spoilage microbial effect on the quality changes of vacuum packed sliced smoked cooked ham
The study of microbial effect on products spoilage was carried out by analysised the spoilage potential of microorganism of its can result in the phenolmenal of spoilage and the spoilage activeity of microorganism of its quantitative ability to produce spoilage metabolites. The results showed that:
Leuconostoc mesenteroides subsp. mesenteroides was indentified as a SSO that contributed to the quality defect such as "shell off", water liquid, "swell bag", relax texture and discolouration, lackluster. It can utilize the glucose of meat medium to heterofermentate to produce CO_2, acetic acid, propylanol, butyricanol, decompose compound of nitrogen such as protein to make the TVB-N value rise up.
Lactobacillus sakei subsp. sakei was identified as a SSO that contributed to quality defects such as sour off-flavor, ropy slime formation. It has preferential fermentation ability to utilize the sugar of meat products to produce lactic acid, acetic acid resulting in pH value drop, exopolysaccharide compound to formate ropy slime, produce representative spoilage compound of tyramine and putrescine resulting in rotten flavour.
Lactobacillus curvatus subsp. melibiosus was identified as a SSO that contributed to the quality defects such as sour off-flavor, ropy slime formation, discoloruration and grayness, rotten flavorur. It has preferential fermentation ability to utilize the glucide of meat products to produce lactic acid, acetic acid and ethanol, and exopolysaccaride compound, furthermore, it can make a great deal of representative spoilage compound of tyramine、cadaverine and putrescine when meat products become spoilage.
4. Analysis of the diversity of microbial community during processing of smoked cooked ham
The diversity of microbial community of the key processing point was analysed by DGGE technique. Through compared the diversity of microbial community between every susceptibly contamination point, the correlation of these microbial community and predominate spoilage bacteria was obtained. The results indicated that:
The main spoilage bacteria of vacuum packed sliced smoked cooked ham are come from the raw meat.
The processing of tumbling and the period of chilling after heat treatment are susceptible contamination points.
Improper using preservative will bring underlying trouble to the product safety.
5. Study of protective effect of bioprotective culture towards the SSO
The effect of Lactobacillus sakei B-2 as bioprotective culture to prolong the shelf-life of sliced vacuum packed smoked cooked ham storaged under 4℃was evaluated. The inhibitory interaction between bioprotective culture and target bacteria was expatiated by characterization of the dynamoic changes of microbial community of meat products during storage under refrigement temperature. The result showed that: The shelf-life of sliced, vacuum packed cooked ham inoculated with L. sakei B-2 at levels of 5.91±0.04 log_(10) CFU/g was 35 days, compared to the 15 days shelf-life of the control product. Treatment with the protective culture has been shown to suppress the growth of the predominant spoilage bacteria including L.sakei, L.curvatus and Le.mesenteroides but can not completely control the growth of strains of uncultured Leuconostoc.
The application of culture-dependent and in-dependent methods led to an understanding of the inhibitory interaction taking place between protective cultures and target strains. PCR-DGGE is an effective technique to analyze the dynamic changes of microorganism population when bioprotective culture applied in pasteurized meat products to prolong the shepf-life. A better understanding of inhibitory interaction between protective cultures and target strains may lead to correct application of protective cultures to meat products.
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