항생제를 이용한 Bifidobacteria의 선택배지

이정준,신명수,나석환,배형석,백영진 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.3

Bifidobacteria가 lactobacilli, streptococci와 혼합되어 있는 유제품으로부터 bifidobacteria의 계수를 위해 bifidobacteria의 선택배지를 개발하였다. 항생제를 포함하는 이 선택배지는 bifidobacteria의 성장에는 영향이 없으면서 다른 유산균의 10^5 cfu/㎖ 이하의 농도를 성장억제시킬 수 있었다. 다른 유산균의 10^5 cfu/㎖ 농도를 성장억제시키기 위해서는 1.0∼2.0 ㎍/㎖의 tetracycline을 BL 한천배지에 첨가해야 하는 것으로 나타났다. Neomycin과 paromomycin을 각각 25 ㎍/㎖의 농도로 1.0 ㎍/㎖의 tetracycline과 혼합하여 BL 한천배지에 첨가할 경우에는, 다른 유산균의 10^6 cfu/㎖ 농도를 성장억제시킬수 있었으나 bifidobacteria의 colony 형성율도 약간 감소하는 것으로 나타났다. 1.0 ㎍/㎖의 tetracycline을 첨가한 BL 한천배지를 bifidobacteria의 선택배지로 사용하면 L. casei, L. acidophilus 그리고 S. salivarius subsp. thermophilus가 bifidobacteria에 비해 1∼10^5배 많은 농도 차이로 존재하는 경우에도 bifidobacteria의 선택적 계수가 가능하였다. Selective agar media were constructed for the counting of bifidobacteria in dairy products containing bifidobacteria, lactobacilli and streptococci. This media containing antibiotics inhibited the growth of lactobacilli and streptococci at less than 10^5 cfu/㎖, but had no influence on the recovery of bifidobacteria. In order to inhibit the growth of 10^5 cfu/㎖ of lactobacilli and streptococci, the addition of 1.0-2.0 ㎍/㎖ of tetracycline in BL agar medium was needed. When 25 ㎍/㎖ of neomycin and paromomycin were mixed with 1.0 ㎍/㎖ of tetracycline in BL agar medium, it was able to inhibit the growth of 10^6 cfu/㎖ of lactobacilli and streptococci but had a little negative effect on the recovery of colonies of bifidobacteria. The results revealed that the BL agar medium containing 1.0 ㎍/㎖ of tetracycline was suitable to count the cell number of bifidobacteria selectively in the presence of a 1 to 10^5-fold excess of L. casei, L. acidophilus and S. salivarius subsp. thermophilus.

유제품으로부터 Bifidobacteria의 선발 및 계수를 위한 선택배지

신명수,이정준,서인영,나석환,백영진 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.2

유제품에서 다른 유산균들과 혼합하여 사용되고 있는 bifidobacteria를 선발하고 계수하기 위하여 두가지 선택배지를 제조하였다. 하나는, phage의 숙주에 대한 감염 특이성을 이용한 것으로서 bifidobacteria와 혼합되어 있는 다른 유산균을 숙주로 하는 phage를 배지에 첨가하여 숙주 유산균을 용균시킴으로써 bifidobacteria에 대한 선택 배지로 사용하는 것이다. 즉, bifidobacteria와 L. casei HY 2782가 혼합된 유제품에서, L. casei HY 2782를 숙주로 하는 J1 phage를 첨가한 BL 배지를 사용하여 bifidobacteria를 계수한 결과, 완전하게 bifidobacteria만을 계수할 수 있었다. 따라서 항생제를 함유하는 선택배지가 아닌 새로운 선택배지로서 그 응용가능성을 제시하였다. 둘째로는 항생제 Tetracycline(1 ㎍/㎖)을 첨가한 선택배지로서 상업적으로 사용되는 bifidobacteria 균주의 성장에는 영향을 주지 않는 반면, Str. salivarius subsp. thermophilus, L. acidophilus, 그리고 L. casei 등의 성장은 억제하였다. 이러한 결과 tetracycline을 사용한 선택배지는 다른 항생제 배지들보다 유제품에 있는 bifidobacteria의 생균수를 정확히 표현해 주었다. Phage utilizing medium and BL agar supplemented with antibiotic Tc(tetracycline) were developed as selective media for the isolation and counting of bifidobacteria in dairy products. The former was based on the host specificity of phage. When bifidobacteria and Lactobacillus casei HY 2782 were mixed together in dairy product, L. casei HY 2782 was lysed by J1 phage which has host specificity to L. casei HY 2782 whereas bifidobacteria grew well on the selective medium added with J1 phage. The latter was found to inhibit the growth of S. salivarius subsp. thermophilus, L. acidophilus, and L. casei, but commercial bifidobacteria grew well in Tc-containing BL agar.

호상 요구르트 제품에 이용되는 Bifidobacteria의 선택적 검출 방법

박희경,허태련 한국산업미생물학회 1995 한국미생물·생명공학회지 Vol.23 No.2

Bifidobacteria를 L. acidophilus, L. bulgaricus와 S. thermophilus 등과 같은 다른 유산균과 함께 제조된 유제품에서 정확한 선택계수를 하기 위해 선택배지에 사용되는 효과적인 항생제와 농도를 조사하고 그 효과를 비교하였다. 본 실험에 사용된 여러 종류의 항생제 중 lithium chloride는 L. acidophilus, L. bulgaricus와 S. thermophilus의 최소 저해농도는 4 ㎎/㎖ 이하이고, B. bifidum, B. longum, B. infantis와 B. breve 균주들은 6∼10 ㎍/㎖로 lithium chloride에 대한 내성이 강하였다. Tetracycline과 chloramphenicol도 다른 유산균에 비하여 bifidobacteria에 최소저해농도가 높아 tetracycline은 1 ㎍/㎖, chloramphenicol은 3 ㎍/㎖를 TPY 배지에 첨가하여 사용하였을 때 L. acidophilus, L. bulgaricus와 S. thermophilus는 저해 효과가 있었으며, bifidobacteria에는 저해 작용이 거의 없었다. Lithium chloride(6 ㎎/㎖), tetrachcline(1 ㎍/㎖)과 chloramphenicol(3 ㎍/㎖) 항생제가 각각 첨가된 TPY 배지는 다른 유산균의 성장 억제를 하고, bifidobacteria에는 억제작용이 거의 없어 선택계수 능이 있었으며, NPNL과 LP 선택배지에 비하여 우수한 선택계수 효과가 있었다. 또한 L. acidophilus, L. bulgaricus, S. thermophilus와 bifidobacteria가 혼합되어 있는 배양액에서 선택계수가 가능하였다. In fermentation of dairy product, bifidobacteria is used in conjunction with other lactic acid bacteria, such as I. acidophilus, L. bulgaricus and S. thermophilus, rendering the enumeration of bifidobacteria difficult. In order to develop optimum conditions for selective enumeration of bifidobacteria, we examined MIC of several antibiotics against various bifidobacteria and other lactic acid bacteria. The growth of L. acidophilus, L. bulgaricus and S. thermophilus were inhibited by lithium chloride at the concentration of less than 4 ㎎/㎖, whereas growth inhibition of bifidobacteria occurred at concentrations over 6∼10 ㎎/㎖. Tetracycline and chloramphenicol were also found to selectively inhibit growth of other lactic acid bacteria at the concentration of 1∼3 ㎍/㎖. Addition of 6 ㎎/㎖ lithium chloride, 1 ㎍/㎖, tetracycline or 3 ㎍/㎖ chloramphenicol to medium was found to be optimal for selective enumeration of bifidobacteria were selectively isolated than with NPNL agar and LP agar.

시판 드링크 및 액상 요구르트 제품중 Lactobacilli 및 Bifidobacteria의 내산성 연구

심명재,이정석,이범진 한국미생물학회 2000 미생물학회지 Vol.36 No.2

국내 시판 19종의 드링크 및 18종의 액상 요구르트 제품에 함유된 2종의 유산균인 Lactobacilli 및 Bifidobacteria의 내산성을 인공위액(pH 1.5) 조건에서 비교 고찰되었다. 드링크 및 액상 요구르트 제품중 초기 Lactobacilli의 생존율은 각각 $10^8~10^{10}$ cfu/ml 및 $10^6~10^{10}$cfu/ml 범위 내에 있었다. 한편 드링크 요구르트 중 초기 Bifidobacteria의 생존율 $10^8~10^{10}$cfu/ml 이었다. 요구르트 제품중 Lactobacilli 및 Bifidobacteria의 생존율은 인공위액에 노출 시간이 증가함에 따라 점차 감소하는 경향을 나타내었으며 120분 노출시에 제품들간에 큰 차이를 나타내었다. (<$10^3~10^6$cfu/ml),캡슐을 함유한 요구르트 제품의 경우, 캡슐을 제외한 Mix는 인공위액에 30분에서 120분 노출시에 약$10^5$cfu/ml이상의 생존율을 나타내었으며 타사 제품과 유사한 거동을 보였다. 그러나 Bifidobacteria를 함유한 캡슐요구르트 제품의 경우 인공위액에 약 120문 노출시켰으때 타사 제품보다 생존율이 약10~737배 이상 증가하였다. 즉 Bifidobacteria의 내산성은 코팅물질을 사용한 캡슐로 봉입하였을 때 증가됨을 알 수 있었다. Gastric acid resistance of aerobic Lactobacilli and anaerobic Bifidohacteria in commercial 19 drink and 18 liquid yogurts in Korea was investigated after exposure to simulated gastric fluid (pH 1.5). The initial survival of Lactobacilli in commercial drink and liquid yogurts was $10^8~10^{10}$ cfulml and $10^6~10^{10}$cfdml, respectively. On the other hand, the initial survival of Bifidobacteria in commercial drink yogurts was ($10^6~10^{10}$cfulml. The survival of Lactobacilli and Bifidobacteria in some commercial drink and liquid yogurts drastically changed depending on the type of commercial products when exposed to simulated gastric fluid for 120 min (<$10^3~10^6$ cfulml). Their survival decreased as a function of time in the simulated gastric fluid. In the case of drink yogurt containing Bifidobacteria-loaded capsu1es;the survival of Bifidobactena in only Mi after excluding capsules was similar to other commercial drink yogurts after exposure to gastric fluid for 30-120 min (ca. TEX>$10^5$</TEX> cfulml). However, the survival of Bifidobacteria in capsules was greatly enhanced due to their stability in low pH condition (>$10^8$ cfulml). The drink yogut containing Bifidobacteria-loaded capsules showed about 10-737 times higher survival in the gastric fluid for 120 min when compared to other commercial drink yogurts. It was evident that the gastric acid resistance of Bifidobacteria could be increased when encapsulated.

${\alpha}-Galactosidase$의 활력차이에 의한 Bifidobacteria의 선별

민해기,이시경,강국희,Min, Hae-Ki,Lee, See-Kyung,Kang, Kook-Hee 한국응용생명화학회 1993 Applied Biological Chemistry (Appl Biol Chem) Vol.36 No.3

본 연구는 합성기질인 $X-{\alpha}-Gal$를 이용하여 발효유 및 유제품내의 Bifidobacteria 생균수를 측정할 목적으로 하였다. 젖산균과 Bifidobacteria의 ${\alpha}-galactosidase$ specific activity를 측정한 결과 Bifidobacteria 균주에서는 높은 ${\alpha}-galactosidase$ activity를 가지고 있었으며, 그 중 Bif. longum KCTC 3215의 specific activity는 8.57 unit/mg protein으로 가장 높게 나타났다. Lactobacillus, Streptococcus, Pediococcus와 Leuconostoc 균주에서는 활성이 미약하거나 없었다. 합성기질인 $X-{\alpha}-Gal$을 MRS agar 배지에 $100{\;}{\mu}M$ 첨가한 결과 Bifidobacteria는 blue colony로, Lac. bulgaricus, Lac. casei와 Leu. mescenteroides 균주는 light blue colony로, 그 외 젖산균에서는 white colony로 나타났다. This method using the synthesis substrate of $5-bromo-4-chloro-3-indolyl-{\alpha}-galactoside\;(X-{\alpha}-Gal)$ was examined for the differential enumeration of Bifidobacteria and lactic acid-producing bacteria. Bifidobacteria possess a high level of ${\alpha}-galactosidase$ activity. Bifidobacterium longum KCTC 3215 exhibited the highest ${\alpha}-galactosidase$ specific activity (8.57 units/mg protein). Determination of ${\alpha}-galactosidase$ activity using the PNPG procedure showed that Lactobacillus, Streptococcus, Pediococcus, and Leuconostoc strain had lower ${\alpha}-galactosidase$ activity as compared to Bifidobacteria. The $X-{\alpha}-Gal$ based medium is useful to identify Bifidobacteria among lactic acid-producing bacteria since the enzyme action of ${\alpha}-galactosidase$ spills $X-{\alpha}-Gal$ substrate and releases indol which impacts a blue color to Bifidobacterial colonies on agar plates. All strains of Bifidobacteria appeared as blue colonies on MRS agar medium supplemented with $100\;{\mu}M\;X-{\alpha}-Gal$ while colonies of other lactic acid-producing bacteria appeared white or light blue.

Comparison of Bifidobacteria Selective Media for the Detection of Bifidobacteria in Korean Commercial Fermented Milk Products

김응률,우건조,김용희,박순옥,조영희,전호남 한국축산식품학회 2010 한국축산식품학회지 Vol.30 No.1

This study was carried out to compare the efficacy and selectivity of TOS and BS media for enumeration of bifidobacteria in commercial fermented milk products. First, bifidobacteria was isolated from 20 fermented milk products, and all isolated bifidobacteria were identified by genomic technology as Bifidobacterium lactis. The two media significantly differed from each other with regard to the recovery of B. lactis, that is, the recovery of this organism was as much as 6 logs lower on BS medium than on TOS. When the concentration of BS solution (mixture of paromomycin sulfate, neomycin, sodium propionate,and lithium chloride) used in BS medium was reduced to 50% (BS50), a relatively high percentage recovery of bifidobacteria from pure cultures was achieved. Susceptibility tests to antibiotics and tests for selective agents for the isolated bifidobacteria and lactic acid bacteria were conducted. The BS solution inhibited some lactic acid bacteria and Bifidobacterium species, while mupirocin (MU) suppressed the growth of all tested lactic acid bacteria but not Bifidobacterium. As compared with BS50 medium, TOS with or without MU showed good bifidobacteria recovery and readily distinguishable colonies; in particular, TOS supplemented with MU had a high selectivity for bifidobacteria. In conclusion, all results suggested that TOS medium with or without MU was found to be suitable for selective enumeration of bifidobacteria from mixed cultures in fermented milk, and better in that capacity than BS medium.

Comparison of Bifidobacteria Selective Media for the Detection of Bifidobacteria in Korean Commercial Fermented Milk Products

Kim, Eung-Ryool,Cho, Young-Hee,Kim, Yong-Hee,Park, Soon-Ok,Woo, Gun-Jo,Chun, Ho-Nam Korean Society for Food Science of Animal Resource 2010 한국축산식품학회지 Vol.30 No.1

This study was carried out to compare the efficacy and selectivity of TOS and BS media for enumeration of bifidobacteria in commercial fermented milk products. First, bifidobacteria was isolated from 20 fermented milk products, and all isolated bifidobacteria were identified by genomic technology as Bifidobacterium lactis. The two media significantly differed from each other with regard to the recovery of B. lactis, that is, the recovery of this organism was as much as 6 logs lower on BS medium than on TOS. When the concentration of BS solution (mixture of paromomycin sulfate, neomycin, sodium propionate, and lithium chloride) used in BS medium was reduced to 50% (BS50), a relatively high percentage recovery of bifidobacteria from pure cultures was achieved. Susceptibility tests to antibiotics and tests for selective agents for the isolated bifidobacteria and lactic acid bacteria were conducted. The BS solution inhibited some lactic acid bacteria and Bifidobacterium species, while mupirocin (MU) suppressed the growth of all tested lactic acid bacteria but not Bifidobacterium. As compared with BS50 medium, TOS with or without MU showed good bifidobacteria recovery and readily distinguishable colonies; in particular, TOS supplemented with MU had a high selectivity for bifidobacteria. In conclusion, all results suggested that TOS medium with or without MU was found to be suitable for selective enumeration of bifidobacteria from mixed cultures in fermented milk, and better in that capacity than BS medium.

Comparison of Bifidobacteria Selective Media for the Detection of Bifidobacteria in Korean Commercial Fermented Milk Products

Eung Ryool Kim,Young Hee Cho,Yong Hee Kim,Soon Ok Park,Gun Jo Woo,Ho Nam Chun 한국축산식품학회 2010 한국축산식품학회지 Vol.30 No.1

This study was carried out to compare the efficacy and selectivity of TOS and BS media for enumeration of bifidobacteria in commercial fermented milk products. First, bifidobacteria was isolated from 20 fermented milk products, and all isolated bifidobacteria were identified by genomic technology as Bifidobacterium lactis. The two media significantly differed from each other with regard to the recovery of B. lactis, that is, the recovery of this organism was as much as 6 logs lower on BS medium than on TOS. When the concentration of BS solution (mixture of paromomycin sulfate, neomycin, sodium propionate, and lithium chloride) used in BS medium was reduced to 50% (BS50), a relatively high percentage recovery of bifidobacteria from pure cultures was achieved. Susceptibility tests to antibiotics and tests for selective agents for the isolated bifidobacteria and lactic acid bacteria were conducted. The BS solution inhibited some lactic acid bacteria and Bifidobacterium species, while mupirocin (MU) suppressed the growth of all tested lactic acid bacteria but not Bifidobacterium. As compared with BS50 medium, TOS with or without MU showed good bifidobacteria recovery and readily distinguishable colonies; in particular, TOS supplemented with MU had a high selectivity for bifidobacteria. In conclusion, all results suggested that TOS medium with or without MU was found to be suitable for selective enumeration of bifidobacteria from mixed cultures in fermented milk, and better in that capacity than BS medium.

Enhanced Delivery of Bifidobacteria and Fecal Changes after Multiple Oral Administrations of Bifidobacteria-loaded Alginate Poly-l-lysine Microparticles in Human Volunteers

Cui, Jing-Hao,Cao, Qing-Ri,Lee, Beom-Jin Informa Healthcare 2007 DRUG DELIVERY Vol.14 No.5

<P> Enhanced delivery of bifidobacteria and fecal changes were compared following multiple oral administrations of protected bifidobacteria-loaded alginate poly-l-lysine microparticles (bap-microparticles) and unprotected bifidobacteria cultures over a period of 1 month to healthy human volunteers as preliminary in vivo studies. When bap-microparticles were orally administered, enhanced delivery of bifidobacteria was achieved. The viability of the bifidobacteria was significantly increased ∼ 11.5-30 times (1.06-1.48 log cycles) during the ingestion period when compared with the bifidobacteria culture group (p < 0.05). However, other gut microflora such as bifidobacteria, enterobacteriaceae, lactic acid bacteria, and staphylococci in feces were not significantly different between the two groups. Encapsulated bifidobacteria resulted in more frequent defecation and decreased fecal viscosity.</P>

시판 요구르트 중 Lactobacilli 및 Bifidobacteria의 안정성 및 내산성 연구

이범진,박옥선,고준수,안태석,박승용 한국미생물학회 1999 미생물학회지 Vol.35 No.1

시판 요구르트 제품중 Lactobacilli 및 Bifidobacteria 의 안정성과 내산성을 비교 고찰하였다. 요구르트 제품중 Lactobacilli 및 Bifidobacteria 의 안정성 및 내산성은 제품들간에 큰 차이가 있음을 고찰하였다. 요구르트 제품중 Lactobacilli 및 Bifidobacteria 의 생존률은 저장중 시간이 지남에 따라 점차 감소하였으나 $10^7$-$10^8$ cfu/ml로의 생존률을 나타내었다. 요구르트 제품중 Lactobacilli 및 Bifidobacteria의 생존률은 pH가 낮아짐에 따라 점차로 감소하였다. 인공위액에서 2시간 노출시킨 후 Lactobacilli 및 Bifidobacteria의 생존률은 3개의 제품에서 $10^3$-$10^4$ cfu/ml 이상이었으나 캡슐제품에 함유된 Bifidobacteria 의 생존률은 $10^8$cfu/ml 이상으로 크게 향상되었다. Stability and gastric acid resistance of Lactobacilli and Bifidobacteria in commercial yogurts were invcstigated. It was noted that there was significant differences of stability and gastric acid resistance among yo-wts. The sutvival of Lactobacilli and B#idohacleria in commercial yogurts decreased as a function of time during storage and showed in the range of $10^7$-$10^8$ cfulml. The lower the pH was, the lower survival of Lactobacillus and B~dobaclerium was observed. The survival of Lactobacillz and Bifidobacteria in three yogurts appeared to be $10^3$-10$^4$ cfuIml. In the case of yogurt containing Bifidobncterza- loaded capsules, the gastric acid resistance of the Rifidobncteria was greatly enhanced and the survival after treatment in a gastric juice for 120 min was over 10' cfulml.