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Injury and Death of Streptococcus lactis due to Freezing and Frozen Storage

机译:冷冻和冷藏导致乳酸链球菌的伤害和死亡

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Cells of Streptococcus lactis were harvested in the early stationary phase, washed, and resuspended in either skim milk (10% nonfat milk) or buffered distilled water (0.0003 m dipotassium phosphate, pH 7.2). Samples of each suspension were frozen and stored at -20 C for intervals up to 28 days. Colony counts of the frozen culture were made using lactic agar and a “restricted” lactic agar medium (Tryptone reduced to 0.5%) to determine injury and death. Death was determined by the difference in plate counts on lactic agar before and after freezing. Injured cells were determined by the difference in plate counts on the two plating media. Greatest injury of the cells occurred during early stages of frozen storage and decreased with time, and death continuously increased. Injury and death were more pronounced when cells were frozen in water than when frozen in 10% nonfat milk solids. Certain cultures survived better when frozen rapidly, whereas with others survival was greater when freezing was slow. Successive freezing, thawing, and propagation of the culture gradually eliminated cells which showed injury by freezing.
机译:在早期固定阶段收获乳酸链球菌细胞,洗涤,然后重悬于脱脂牛奶(10%脱脂牛奶)或缓冲蒸馏水(0.0003 m磷酸氢二钾,pH 7.2)中。将每种悬浮液的样品冷冻并在-20 C下保存28天。使用乳酸琼脂和“限制性”乳酸琼脂培养基(色氨酸降至0.5%)对冷冻培养物进行菌落计数,以确定损伤和死亡。通过冷冻前后乳酸琼脂平板计数的差异来确定死亡。通过两种平板培养基上平板计数的差异来确定受伤的细胞。细胞的最大损伤发生在冷冻储存的早期阶段,并随时间减少,并且死亡持续增加。当将细胞在水中冷冻时,与在10%脱脂牛奶固体中冷冻相比,损伤和死亡更为明显。快速冷冻时,某些培养物的存活率更高,而缓慢冷冻时,其他培养物的存活率更高。培养物的连续冷冻,解冻和繁殖逐渐消除了由于冷冻而显示出损伤的细胞。

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