The cytidine deaminase was partialy purified with sephadex G-200 and the characteristics of the enzyme were clarified. The molecular mass of the plasmid-encoded protein was identified as about 30 kDa in a minicell system. The native enzyme was estimated t
The Salmonella typhimurium cdd gene encoding cytidine deaminase (cyti-dine/2'-deoxycytidine aminohydrolase; EC 3.5.4.5.) was isolated through shotgun clon-ing by complementation of the E. coli odd mutation. By subsequent deletion and sub-cloning from the
동치미액은 전통적으로 냉면국물로 이용되어왔다. 냉면국물의 미생물 오염문제를 해결하기 위하여 항균활성이 높은 김치 젖산균 Lactobacillus homohiochii B21과 Leuconostoc mesenteroides subsp. mesenteroides C16을 스타터로 혼합사용하여 동치미액을 제조하고, 이를 냉면국물에 100%, 50%, 10% 및 0% 첨가하여 20℃ 및 10℃에서 보관할 때에 동치미액의 항균력으로 인하여 의도적으로 첨가한 Salmonella typhimurium과 Staphylococcus aureus의 증식이 억제되는 정도를 조사하였다. 동치미액 100%인 냉면국물은 20℃에서 보관할 때는 Salmonella typhimurium은 16시간만에, Staphylococcus aureus는 32시간만에 10^6CFU/ml에서 10^0CFU/ml로 각각 급격히 사멸하였고, 10℃에서 보관할 때도 시간경과에 따라 생균수가 급격히 감소하였으나 감소속도는 20℃의 경우보다 느렸다. 동치미액을 50% 첨가한 육수에서도 Salmonella typhimurium과 Staphylococcus aureus는 시간경과에 따라 급격히 감소하였으며, 감소속도는 동치미액 100%일 경우보다 느렸다. 동치미액을 10% 첨가한 육수에서는 20℃에서 보관할 때는 초기에 Salmonella typhimurium과 Staphylococcus aureus의 균수증가가 약간 있었으나 동치미액 무첨가에 비하여 현저히 억제되었으며, 8시간∼16시간 이후부터는 균수가 서서히 감소하였다. 육수에 동치미액을 첨가하지 않았을 경우에는 20℃에서 보관할 때는 초기부터 Salmonella typhimurium과 Staphylococcus aureus의 균수증가가 급격히 이루어졌으며, 10℃에서 보관할 때는 24시간 이후부터 서서히 증가하였다. Salmonella typhimurium과 Staphylococcus aureus에 대한 본 동치미액의 억제력은 후자의 미생물보다 전자에 대하여 더 강하게 나타났다. 본 연구의 동치미액을 육수에 50% 첨가하여 냉면국물을 제조한다면 보관중의 미생물 오염문제 해결에 큰 도움이 될 것으로 판단된다.
This study was performed to investigate the changes of amount of S. typhimurium during cooking processes using pork and japchae (a Korean food which is made from meat, vegetables and noodles), and to support a practical application to develop a hazard analysis critical control point (HACCP) model. The pork was purchased in a retail shop, cut (0.5 cm × 10 cm × 10 cm, 25 g), tested for Salmonella contamination (results : negative), inoculated with S. typhimurium (10^7 CFU/g), then treated in various conditions related to cooking. After thawing for 24 hours in various conditions, the number of S. typhimurium was increased to 10^(10) CFU/g at a refrigerated temperature (4-10℃), and to 10^(21) CFU/g at room temperature (22-29℃). After thawing in a microwave oven for 40 seconds, the number of S. typhimurium increased to l0^8 CFU/g. During the thawing period, the number of S. typhimurium increased over time. At the refrigerated temperature, the number of the bacteria was 10^(10) CFU/g after 24 hours, 10^(13) CFU/g after 48 hours, and 10^(20) CFU/g after 72 hours. At room temperature the number of bacteria reached 10^(11) CFU/g in 2 hours, 10^(15) CFU/g in 4 hours, 10^(16) CFU/g in 8 hours, 10^(18) CFU/ g in 12 hours, and 10^(21) CFU/g in 24 hours. After cooking in a frying pan (150±7℃) for 3 minutes, the bacterial count was 10^6 CFU/g. After cooking in hot water for 20 minutes, the bacterial count was 10^7 CFU/g at 60℃, 10^6 CFU/g at 63℃, and 10⁴ CFU/g at 65℃. The fried pork was mi×ed with cooked vegetables, noodles, sesame oil, sesame seeds, and seasonings to make Korean japchae. This process took 10±2 minutes. The bacterial count in the japchae increased to 10^7 CFU/g from the count of 10^6 CFU/g of the fried pork before it was mixed with the other ingredients. These results indicate that the amount of S. typhimurium is effected by various different cooking processes. This study can suggest that pork should be cooked in water at over 65℃ for 20 minutes in order to prevent food poisoning, if the pork is contaminated with S. typhimurium. The presence of S. typhimurium in the raw pork is identified in an HA for japchae, and the primary CCP for japchae is inadequate cooking (cooking method and time/temperature). We need to standardize time-temperature-size and amount of pork in cooking japchae, because pork is usually cooked in ordinary frying pans when we make this food.
Lactobacillus acidophilus NCFM, Lactobacillus casei YIT 9018, Bifidobacterium longum 8001, and Bifadobacterium longum 8025 at the level of 10^6 cfu/ml were cultured with 10⁴ cfu/ml of Escherichia colt O157:H7 KSC 109 or Salmonella typhimurium ATCC 14028, in order to verify the effects of lactic acid bacteria and bifidobacteria on the growth of the pathogens. In the mixed culture of lactic acid bacteria with E. colt O157:H7 KSC 109, growth inhibition and atypical microcolonies of E. colt O157:H7 KSC 109 were observed. The pathogens inoculated grew for 5 hours (pH 5.3), by the time L. acidophilus NCFM reached the exponential growth phase, and then the surviving pathogens were decreased to 10¹ cfu/ml after 35 hours. When L. casei YIT 9018 was grown with the pathogens, they grew for 10 hours (pH 4.6), by the time L. casei YIT 9018 reached the end of exponential growth phase, and then the surviving pathogens were decreased drastically. Up to the stationary growth phase of lactic acid bacteria, L. acidophilus NCFM exhibited stronger inhibition against the pathogens than L. casei YIT 9018 did, which might be attributed to its faster growth. Likewise bifidobacteria inhibited the growth of the pathogens. tested, bifidobaceria was weaker in the inhibitory activity than lactic acid bacteria. When Bifidobacterium longum 8001 was cultured with the pathogens, E. colt O157:H7 KSC 109 was gradually inhibited at the stationary growth phase of bifidobacteria, atypical microcolonies were formed on Levine EMB medium after 48 hours, and Salmonella grew up to 10^6 cfu/ml, then was drastically inhibited at the exponential growth phage of Bifxdobacterium longum 8001. But when Bifidobacteriuam longum 8025 was cultured with the pathogens, the pathogens grew to the same level of Bifidobacteriuam longum 8025 after 10 hours, then the surviving pathogens were decreased drastically.
Escherichia coli O157:H7 and Salmonella ser. typhimurium are pathogens involved in food poisoning in numerous countries. This study aimed to obtain knowleges on the survival of E. coli O157:H7 KSC 109 and S. ser. typhimurium ATCC 14028 in fermented milk products which were on sale in Suwon Yakult supplier. To the final concentration of 10³-10⁴ cfu/ml of E. coli O157:H7 KSC 109 or S. ser. typhimurium ATCC 14028 in the fermented milks, Metchnikoff, Ace, Yakult, Mastoni and Super100 were inoculated with these pathogens and then were stored at 4 and viable cells of these pathogens were periodically counted. The results showed that the survival of two pathogens differed in the different types of fermented milks tested. Number of surviving E. coli O157:H7 KSC 109 and S. ser. typimurium ATCC 14028 cells (initial inoculum, 10³-10⁴ cfu/ml) were decreased to 10', 102 cfu/ml in Ace after 100 hours, and were decreased gradually to 10¹ cfu/ml in Yakult after 250 hours. In the other fermented milks, viable cells of E. coli O157:H7 KSC 109 was not drastically decreased but those of S. ser. typhimurium ATCC 14028 was decreased gradually to 10² (Mastoni), and to 10¹ cfir/ml (Super100) after 250 hours. It appeared that S. ser. typhimurium ATCC 14028 was more susceptible than E. coli O157: H7 KSC 109 at low pH. Vibale cells of E. coli O157:H7 KSC 109 was not drastically decreased in most of fermented milks tested except Ace and Yakult, but in general, S. ser. typhimurxum ATCC 14028 was drastically decreased in most of the fermented milks. The major inhibition factor against these pathogens in the fermented milks during storage at 4℃ appeared to be the acidity and the metabolites produced by the starters bacteria used in fermented milk products.
The growth inhibition effect of orally administrated yogurt ACE and Metchnikoff upon E. coli O157:H7 and S. typhimurium inoculated into gastric lumen of rabbits was investigated. The rabbits challenged with each 1 ml of suspension containing 10^8 CFU/ml of the pathogens were divided into 4 groups by the interval of yogurt administration: A group; preadministrated 7 days before inoculation of the pathogens and fed daily; B group; administrated daily after inoculation of the pathogens, C group; administrated every 3 days after inoculation of the pathogens; Control group, not fed after inoculation of the pathogens. Each 3 ml of yogurt containing 10^9 CFU/ml was orally administrated into rabbits. All yogurt administrated groups (A, B, C) showed growth inhibition effect on E. coli O157:H7 in one day after inoculation of the pathogen by the level of 0.8-1.0 log CFU/g, compared with the result differences between the control group and the yogurt administrated groups. In the control group after 5 days of inoculation, the number of colonized pathogens was 10^5-10^6 CFU/g, whereas 10³-10⁴ CFU/g was detected in the yogurt administrated groups. After 10 days of inoculation, the viable pathogen number per gram (g) of the rabbit feces was 10³ CFU/g in the control group, whereas the number below 10¹ CFU/g was detected in the group A, and 10² CFU/g in the group B and C. The growth inhibition effect of yogurt administration on E. coli O157:H7 was highly increased in the order of A, B, and C group. The same effect on S. typhimurium was observed at the level of 2 log CFU/g in the Metchnikoff yogurt administrated groups, compared with the control group result in one day after inoculation of the pathogen. In 7 days after inoculation of the pathogen, the viable number was increasingly decreased, and finally after 15 days no viable cell of S. typhimurium was discharged into the fecal samples in the group A, and the mean level of 10¹ CFU/g was detected in the group B, but there was no growth inhibition effect in the group C. The growth inhibition effect on S. typhimurium was observed at the same level of viable cell number between the yogurt ACE administrated groups and the control group in 5 days after inoculation. But, after 10 days of inoculation the viable cell number was started to decrease, and the viable cell of S. typhimurium was not discharged from rabbit intestinal contents after 15 days of inoculation in the yogurt ACE administrated groups. In such a case that yogurt was administrated in order to prevent the pathogens, pre-administration on a daily basis one week before inoculation of the pathogens exerted considerable effect in growth inhibition. In comparison with two kinds of yogurt tested in this study, the growth inhibition effect on two kinds of pathogens was observed more highly in the Metchnikoff administrated group than the ACE administrated group.
새싹 채소 중 무순의 위생적인 재배 조건을 설정하기 위하여 종자세척 및 관수로서 이산화염소수, citric acid 및 glycerin의 적용 효과를 조사하였다. 무 종자에 인위적으로 오염시키기 위하여 Salmonella typhimurium은 실험실 조건에서 배양하였으며, 무 종자를 S. typhimurium 현탁액에 침지시켜 균수를 약 로 조절하였다. 무 종자에 대한 세척 효과는 이산화염소수 및 citric acid의 단독처리보다 병행처리시
Water extract of green tea(GTW) and 70% ethanol extract of green tea(GTE) were prepared for the test of antibacterial activity. The sensitivity of Staphylococcus aureus and Salmonella typhimurium to the green tea extracts in various pH of culture broth was tested. Tryptic soy broth(TSB) containing 0∼2%(w/v) of green tea extracts was adjusted to pH 5.0∼7.0 and inoculated with 105/∼106/ cells/ml of each bacteria. The plate counting method and clear zone test were used to determine inhibitory effect of green tea extracts. Green tea extracts completely inhibited the growth of S. aureus at 0.5% level and bactercidal at 0.5∼1.0% level of GTW and GTE at pH 5.0∼7.0. Green tea extracts were bactercidal to S. typhimurium at 1.5∼2.0% level of GTW and 1.0∼2.0% level of GTE at pH 7.0. Sal. typhimurium was more resistant than S. aureus. in same concentration of green tea extracts at same pH. The resistance of S. aureus and Sal. typhimurium was increased with decreasing pH of culture broth. The morphology of S. aureus cells treated with green tea extracts showed damage of cell wall and cytoplasmic membrane. Severely damaged cells of S. aureus lost electron dense material and cytoplasm. Green tea extracts stimulated autolysis and cell death of S. aureus. This result suggests that green tea extracts can be used as an effective natural antibacterial agent in food.