• Title/Summary/Keyword: enrichment broth

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A Comparison of three Enrichment Media for Isolating Salmonella (Salmonella균(菌) 분리용(分離用) 증균배지(增菌培地)의 비교실험(比較實驗))

  • Kim, Yong-Ja;Lee, Seung-Yun;Park, Kee-Deuk;Min, Chang-Hong
    • The Journal of the Korean Society for Microbiology
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    • v.11 no.1
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    • pp.33-48
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    • 1976
  • The practical significance of using a selective enrichment procedure for detecion and enumeration of salmonella is well recognized. There are still various selective enrichment media has been communly used. Early years selenite broth was recomnended as an enrichment media for the isolating of salmonella. Hajna introduced a modified tetrathionate broth and demonstrated the greater efficiency to compare with the previous enrichment media. Raj also described that the new medium called dulcitol selenite enrichment and has been found to be very satisfactory, especially general implication in food poisoning. Authors tried to compare these 3 enrichment media for isolating salmonella. 1. When salmonella strains were inoculated $1{\sim}10^6$ cells per tube to these 3 enrichment media, mostly similar results were obtained between selenite broth and DS broth. In these 2 enrichment broth were showed $10^7/ml-10^8/ml$ cells of all tested salmonella strains. But in the case of TT broth it was found that the growth was $10^3/ml{\sim}10^4/ml$ cells for tested strain. 2. When E. coli, Proteus, Citrobacter were inoculate $10{\sim}10^6$ cells per tube to these 3 enrichment media. It was suggested that DS broth was showed more inhibitory action than that of selenite broth. TT broth showed high inhibition to these 3 organisms tested. 3. It was generally known that the incubation time is influenced to the frequency of salmonella detection. For this tendency, DS broth and selenite broth were showed similar results within 24 hrs to 48hrs incubation to the test. But DS broth showed more inhibitory action to E. coli and Proteus than that of selenite broth. 4. When $1{\sim}10$ cells were inoculated(per tube) to these 3 enrichment media, DS broth was found to be more sensitive than that of selenite broth.

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Comparison of Enrichment Media of Shigella sonnei (쉬겔라 증균배지의 성능 비교)

  • In, Ye-Won;Ha, Su-Jeong;Kim, Seok-Joong;Oh, Se-Wook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.12
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    • pp.1787-1792
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    • 2011
  • The object of this study was to compare the performance of commercial enrichment media used for Shigella spp. A total of four enrichment media, Gram negative (GN) broth, Shigella broth (SB), selenite-F (SF) broth, and selenite cystine (SC) broth, were tested. When S. sonnei was inoculated into each enrichment broth at 10 cfu/mL of concentration, the highest growth was observed in Shigella broth. Morganella spp., which was not differentiated in selective agar of Shigella spp. thus can be counted as false positive, did not grow in Shigella broth in enrichment step. When S. sonnei was artificially inoculated into pork, it was mostly recovered through an enrichment process with GN broth and SF broth. However, in the case of beef, S. sonnei was mostly recovered with GN broth but largely failed with Shigella broth. Therefore, enrichment media for Shigella spp. should be selected by considering the food matrix in order to increase the chance of isolating it from foods.

Determination of best enrichment media for growth of Salmonella injured from cold temperature during process and storage (저온저장으로 인해 손상된 살모넬라를 배양하기 위한 최적의 배지 선정에 관한 연구)

  • Park, Mi-Kyung
    • Food Science and Preservation
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    • v.23 no.6
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    • pp.759-764
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    • 2016
  • This purpose of this study was to determine the best enrichment medium for rejuvenating and recovering Salmonella placed in cold temperature prior to the employment of the gold biosensor combined with a light microscopic imaging system. A mixture of nalidixic-resistant Salmonella Typhimurium and Enteritidis were inoculated onto chicken (1,000 CFU/chicken). After cold injury at $4^{\circ}C$ for 24 hr, Salmonella on chicken was enriched for 6 hr with six non-selective media including buffered peptone water broth, lactose broth, brain heart infusion broth (BHI), universal pre-enrichment broth, nutrient broth, and tryptic soy broth, and five selective media including brilliant green broth (BG), rappaport-vassiliadis R10 broth, selenite cystine broth, selenite broth, and tetrathionate brilliant green broth (TBG) for the comparison of Salmonella growth. Various concentrations of Salmonella (10, 50, 100, 500, and 1,000 CFU/chicken) were then enriched for 6 hr in both BHI and BG media to select the best media. BHI was selected as the most effective non-selective enrichment medium, while BG was selected as the most effective selective enrichment medium. Finally, BHI medium was selected as the most efficient enrichment medium for Salmonella growth injured from cold temperature during processing or storage.

Salmonella sp. 의 신속한 동정을 위한 증진배양의 개선에 관한 연구

  • Kim, Kee-Tae;Kim, Tae-Ue;Yook, Soon-Hak;Pek, Un-Hua
    • Microbiology and Biotechnology Letters
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    • v.24 no.6
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    • pp.647-651
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    • 1996
  • The development of an enrichment method for the rapid and effective identification of Salmonella spp. in sewage or food was studied. As a growth factor for Salmonella, 10 mM cyclic adenosine monophosphate (cAMP) in trypticase soy broth with 0.6% yeast extract (TSBYE) increased cell number five-folds and 0.6% yeast extract in selenite broth increased cell number ten-folds of control. Bile salts in selenite broth was tested for the selection of S. enteritidis in a mixture with Staphylococcus aureus, Pseudomonas aeruginosa, Lactobacillus plantarum and Escherichia coli. The latter four strains were effectively inhibited at 0.1% bile salt. A two-step culture method was used to enrich Salmonella spp.; a primary-enrichment and secondary- enrichment culture. At a primary-enrichment step, selenite broth with 0.6% yeast extract and 10 mM cAMP was used, and at a secondary-enrichment step, 0.1% bile salt was additionally used. Culture times of a primary- enrichment and a secondary-enrichment step were 8 hr and 6 hr, respectively. In this procedure, cell number increased from 10$^{0.3}$ to 10$^{8.5}$ with inhibition of other strains within 14 hr. In the case of an initial cell concentrarion as low as 10$^{-2}$ cfu/ml, a cell number increased to 10$^{7}$ cfu/ml by using a 10 hr primary-enrichment and 6 hr secondary-enrichment procedure.

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Studies on the Methods for the Isolation of salmonella from Pigs (돼지에서 살모넬라균(菌)을 분리(分離)하는 방법(方法)에 관한 연구(硏究))

  • Chung, G.T.
    • Korean Journal of Veterinary Research
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    • v.9 no.1
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    • pp.63-69
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    • 1969
  • Various methods for the isolation of salmonellae were compared by examining mesenteric lymph nodes and faeces of 395 pigs at slaughter. The conclusions were as follows. (i) Tetrathionate broth was significantly superior to selenite broth for the examination of mesenteric lymph nodes. (ii) There was no significant difference between tetrathionate broth and selenite broth for the examination of faeces. The use of both tetrathionate broth and selenite broth as enrichment media for faeces produced far better results than the use of any single enrichment medium. (iii) Brilliant green agar was significantly superior to SS agar for the examination of mesenteric lymph nodes. (iv) There was no significant difference between brilliant green agar and SS agar when faeces were examined. The use of both media produced far better results than the use of either singly. (v) Brilliant green MacConkey broth was much inferior to tetrathionate broth as an enrichment broth. Direct culture of faeces on brilliant green MacConkey agar yielded less isolates than prior enrichment in tetrathionate or selenite broth. (vi) The optimum incubation period of enrichment was 24 hours for faeces and 48 hours for mesenteric lymph nodes when either tetrathionate broth or selenite broth was used as enrichment media.

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Comparison of 10 Different Pre-Enrichment Broths for the Regeneration of Cronobacter spp. (Enterobacter sakazakii ) Infected in Powdered Infant Formula

  • Jung-Whan Chon;Kun-Ho Seo;Hyungsuk Oh;Dongkwan Jeong;Kwang-Young Song
    • Journal of Dairy Science and Biotechnology
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    • v.41 no.3
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    • pp.103-112
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    • 2023
  • This study aimed to assess the effectiveness of 10 different pre-enrichment methods using Real-Time polymerase chain reaction (PCR) in support of the FDA method. When the initial Cronobacter spp. (Enterobacter sakazakii) inoculation was 7.2 CFU/g, the Ct values were observed in the following order: 21.37 (Enterobacteriaceae enrichment [EE] broth), 21.95 (brain heart infusion [BHI]), 22.72 (tryptic soy broth [TSB]), 23.02 (violet red bile lactose [VRBL]), 22.31 (TSB-0.1% sodium pyruvate [SP]), 23.43 (distilled water [DW]), 24.34 (phosphate buffered saline [PBS]), 24.95 (nutrient broth [NB]), 25.82 (TSB-0.6% yeast extract [YE]), and 28.27 (violet red bile glucose [VRBG]). For an inoculation of 1.82% CFU/g of Cronobacter spp. (E. sakazakii), the Ct values were recorded in this sequence: 20.34 (EE broth), 22.16 (TSB-0.6% YE), 22.37 (BHI), 22.71 (VRBL), 22.88 (TSB), 23.01 (DW), 23.19 (NB), 23.79 (TSB-0.1% SP), 24.66 (VRBG), and 24.70 (PBS). Finally, when the inoculum of Cronobacter spp. (E. sakazakii) was 0.182 CFU/g, the Ct values followed this order: 21.93 (VRBL), 23.07 (TSB-0.6% YE), 23.31 (DW), 23.47 (PBS), 23.70 (BHI), 24.14 (TSB-0.1% SP), 25.14 (TSB), 29.00 (VRBG), 31.55 (EE broth), and were undetected in the case of NB. Consequently, these results indicate that there were no significant differences among the 10 different pre-enrichment broths. Future studies should focus on exploring pre-enrichment broths that can improve the limit of detection at very low Cronobacter spp. (E. sakazakii) concentrations and enhance the selective recovery of Cronobacter spp. (E. sakazakii) under acid, antibiotic, cold, and heat damage conditions.

Comparison of Real-Time PCR and Conventional Culture Method for Detection of Cronobacter spp. in Powdered Foods (분말식품에서 Cronobacter spp. 검출을 위한 Real-Time PCR과 배지배양법의 비교검증)

  • Chon, Jung-Whan;Song, Kwang-Young;Kim, Sun-Young;Hyeon, Ji-Yeon;Kim, Yun-Gyeong;Hwang, In-Gyun;Kwak, Hyo-Sun;Seo, Kun-Ho
    • Korean Journal of Microbiology
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    • v.47 no.1
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    • pp.87-91
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    • 2011
  • The aim of this study was to compare the performance of conventional culture and real-time PCR for detection of Cronobacter spp. in powdered foods. Infant formula, baby food and Misugaru inoculated with Cronobacter were enriched in distilled water as first enrichment step, followed by incubating in Enterobacteriaceae enrichment (EE) broth as second enrichment step. A loopful of enriched sample was streaked onto Druggan-Forsythe-Iversen agar, followed by incubating at $37^{\circ}C$ for 24 h. One milliliter of the enriched distilled water and EE broth were used in real-time PCR assay. No statistical differences were observed in the number of positive samples between culture method and real-time PCR (p>0.05) in all types of food samples. The number of positives of real-time PCR was higher in the first enrichment media (distilled water) than the second enrichment media (EE broth), though there was no significant difference (p>0.05). It appears that some components of the second enrichment broth, EE broth, inhibit the reaction of real-time PCR. These results show that real-time PCR using a single enrichment with distilled water could be useful as an effective screening method for detection of Cronobacter while saving much time and labor compared to conventional culture method.

Evaluation of enrichment broth and selective media for the detection of non-O157 enterohemorrhagic Escherichia coli (Non-O157 장출혈성대장균 검출을 위한 증균배지 및 선택배지 성능 평가)

  • Lee, Da Yeon;Kim, Hee-eon;Seo, Dong Won;Cho, Yong Sun
    • Korean Journal of Food Science and Technology
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    • v.48 no.4
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    • pp.323-328
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    • 2016
  • In this study, specific and rapid enrichment and growth conditions for the most important, classic non-O157 enterohemorrhagic Escherichia coli (EHEC) serogroups were assessed. Three enrichment broth types, namely, EC medium with MUG broth, BRILA broth, and mTSB broth with novobiocin, were compared to identify the optimum enrichment broth for EHEC isolation. Four kinds of selective media, namely, ENDO agar, Chromocult agar, TBX agar, and CHROMagar$^{TM}$ STEC medium, were compared to identify the optimum one for non-O157 EHEC isolation. The results suggested that incubation in EC medium with MUG broth at $42^{\circ}C$ for 20 h is optimum for the enrichment of non-O157 EHEC. TBX agar was identified to have the highest specificity among the tested media. Consequently, a combination of complementary selective media according to serotype must be considered for comprehensive isolation of specific EHEC.

Evaluation of the Selective Enrichment Culture to Recover Clostridium difficile

  • An, Byoungrak;Kim, Heejung;Lee, Kyungwon
    • Korean Journal of Clinical Laboratory Science
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    • v.46 no.4
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    • pp.140-142
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    • 2014
  • To evaluate the recovery rates to increase toxigenic C. difficile, the selective enrichment broth culture methods were compared with commonly used cytotoxin assays and toxigenic culture. First, the enrichment culture, using the selective medium broth for 2 to 5 days, was performed and then, toxigenic C. difficile was confirmed by C. difficile toxin gene-specific PCR after being cultured on C. difficile selective agar. The sensitivity of C. difficile from the enrichment culture (100%) was higher than that of C. difficile selective agar culture (93.8%), while positive predictive values (PPV) were low; 72.7% (16/22) and 88.2% (15/17). PPV of the enrichment culture are not high. Recently, combinations of C. difficile selective agar culture, C. difficile A & B assays, glutamate dehydrogenase, and nucleic acid amplification method are widely used. The enrichment culture was disadvantageous in PPV, turn-around time, and cost. So, what we performed is not considered as a common method of diagnosis of C. difficile-associated diarrhea.

Experimental Studies on Detection of Salmonellae in Animal-origin Foods by Means of Dirct Fluorescent Antibody Technique (직접형광항체법(直接螢光抗體法)에 의한 축산식품중(畜産食品中)의 Salmonella 균(菌) 검출(檢出)에 관한 실험적(實驗的) 연구(硏究))

  • Jeon, Moo Hyeong;Cha, Youn Ho;Chung, Gill Taik
    • Korean Journal of Veterinary Research
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    • v.14 no.2
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    • pp.243-252
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    • 1974
  • The experiment was performed in order to investigate the applicability of the rapid detection of salmonellae in various animal-origin foods by means of the direct fluorescent antibody technique. Egg, sausage and chicken were inoculated with various concentrations of Sal.paratuphi A, Sal. paratyhi B and Sal. thompson, and the fluorescent antibody technique was applied and compared with the conventional cultura method for the sensitivity of detection of the organisms. Two methods were employed in the fluorescent antibody technique; the direct smear method in which the smear being made directly from the specimens, and the enrichment smear method in which the smear being made from the enrichment broth. The effect of various enrichment time (1,5,8,11 and 13 hours) in tetrathionate broth on the detection of salmonellae in the fluoresent antibody technique was also studied. The results obtained were summarized as followings; 1. Of the three methods, the enrichment smear method of fluorescedt antibody technique was highly effective as cultural method for the detection of salmonella organisms. 2. Direct smear method of fluorescent antibody technique was effective as two other methods $5{\times}10^4$ organisms presented in 50 g(ml) of specimens. This method may not be applicable when the specimens contained $5{\times}10^2$ or less organisms. 3. Of the three specimens, the recovery rate of Salmonella organisms from egg was slightly higher than that of sausage and chicken. 4. In fluorescent antibody technique and cultural method, the specimens inoculated with Sal. thompson were found to be higher detection rate than the specimens inoculated with Sal. paratyphi A, 5. The optimum enrichment time of Salmonella organisms in tetrathionate broth on the detection by fluorscent antibody technique was found to be 11 hours or longer when the specimens of egg, sausage and chicken were inoculated with approximately 500 organisms. The longer enrichment time was the higher detection rate up to 11 hours tested.

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