• Asian-Australasian Journal of Animal Sciences
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• 제20권12호
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• pp.1887-1894
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• 2007

In this paper, a rapid and reliable gene-targeted species-specific polymerase chain reaction (PCR) technique based on a two-step process was established to identify bifidobacteria in dairy products. The first step was the PCR assay for genus Bifidobacterium with genus specific primers followed by the second step, which identified the species level with species-specific primer mixtures. Ten specific primer pairs, designed from nucleotide sequences of the 16-23S rRNA region, were developed for the Bifidobacterium species including B. angulatum, B. animalis, B. bifidum, B. breve, B. catenulatum, B. infantis, B. longum, B. minimum, B. subtile, and B. thermophilum. This technique was applied to the identification of Bifidobacterium species isolated from 6 probiotic products, and four different Bifidobacterium spp. (B. bifidum, B. longum, B. infantis, and B. breve) were identified. The findings indicated that the 16S-23S rDNA gene-targeted species-specific PCR technique is a simple and reliable method for identification of bifidobacteria in probiotic products. PCR combined with Denaturing Gradient Gel Electrophoresis (DGGE) for identification of the bifidobacteria was also evaluated and compared with the gene-targeted species-specific technique. Results indicated that for fermented milk products consistency was found for both species-specific PCR and PCR-DGGE in detecting species. However, in some lyophilized products, the bands corresponding to these species were not visualized in the DGGE profile but the specific PCR gave a positive result.

• Journal of Microbiology and Biotechnology
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• 제30권6호
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• pp.885-892
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• 2020

Chemotherapy regimens for non-small cell lung cancer (NSCLC) have various adverse effects on the human body. For this reason, probiotics have received attention regarding their potential value as a safe and natural complementary strategy for cancer prevention. This study analyzed the anticancer effects of aqueous extracts of probiotic bacteria Bifidobacterium bifidum (BB), Bifidobacterium longum (BL), Bifidobacterium lactis (BLA), Bifidobacterium infantis 1 (BI1), and Bifidobacterium infantis 2 (BI2) on NSCLC cell lines. When the aqueous extracts of probiotic Bifidobacterium species were applied to the NSCLC cell lines A549, H1299, and HCC827, cell death increased considerably; in particular, the aqueous extracts from BB and BLA markedly reduced cell proliferation. p38 phosphorylation induced by BB aqueous extract increased the expression of cleaved caspase 3 and cleaved poly (ADP-ribose) polymerase (PARP), consequently inducing the apoptosis of A549 and H1299 cells. When the p38 inhibitor SB203580 was applied, phosphorylation of p38 decreased, and the expression of cleaved caspase 3 and cleaved PARP was also inhibited, resulting in a reduction of cell death. In addition, BB aqueous extracts reduced the secretion of MMP-9, leading to inhibition of cancer cell invasion. By contrast, after transfection of short hairpin RNA shMMP-9 (for a knockdown of MMP-9) into cancer cells, BB aqueous extracts treatment failed to suppress the cancer cell invasiveness. According to our results about their anticancer effects on NSCLC, probiotics consisting of Bifidobacterium species may be useful as adjunctive anticancer treatment in the future.

• Journal of Microbiology and Biotechnology
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• 제15권2호
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• pp.388-394
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• 2005

In order to investigate the distribution of dominant Bifidobacterium species in intestinal microflora of Korean adults and seniors, SDS-PAGE profiles of whole cell proteins were used for the identification of bifidobacteria. To confirm the reliability of SDS-PAGE, the Bifidobacterium species identified by SDS-PAGE of whole cell proteins were validated by using 16S rDNA sequencing analysis. The results of SDS­PAGE corresponded well with those determined by the analysis of 16S rDNA sequencing. Based on the analysis of SDS-PAGE patterns on unidentified fecal strains which showed positive in fructose-6-phosphate phosphoketolase activity, B. adolescentis, B. longum, and B. bifidum were identified in the feces of adults, and B. adolescentis, B. longum, B. bifidum, B. breve, and B. dentium were identified in those of seniors. In most of the fecal samples tested, the predominant Bifidobacterium species consisted of only a few species, and differences in the distribution and numbers of Bifidobacterium species were observed between adults and seniors. B. adolescentis and B. longum were found to be the most common species in feces of adults, but not in seniors. Accordingly, the distribution and abundance of bifidobacteria in the human intestinal microflora varied depending on the age of hosts.

• Journal of Microbiology and Biotechnology
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• 제12권1호
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• pp.176-181
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• 2002

In order to isolate and identify Bifidobacterium spp. that originated in Korea, feces were sampled from healthy Korean adults and children living in three villages, the first having a history of longevity and the other two where the diet did not include fermented milk or any pharmaceutical preparations. Through the use of Gram staining and microscopic examination for cell morphology, 23 bacterial strains presumed to be the Bifidobacterium genus were isolated from the feces of 13 out of a total of 59 Korean people. To identify the Bifidobacterium strains at the genus level, these bacteria were then analyzed by TLC and the fructose-6-phosphate phosphoketolase (F6PPK) test. The result showed that 22 of the isolated strains were confirmed to be members of the genus Bifidobacterium. All of these bifidobacteria were also identified as Bifidobacterium spp. by the fermentation test. Using a RFLP analysis, an attempt was made to identify the Bifidobacterium spp. that had been isolated from both Korean adults and children. In a genomic Southern blot analysis after digestion with two restriction enzymes (EcoRI, HindIII), all of the 14 randomly selected Korean isolates showed patterns identical to those of three different B. longum species. Another restriction enzyme, CfoI (4-bp recognition enzyme), was then used to identify the strain. Interestingly, all the Korean isolates were identified as B. longum ATCC 15708, indicating that a RFLP analysis was effective for identifying Bifidobacterium spp. at both the strain and species levels.

• 약학회지
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• 제48권1호
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• pp.20-26
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• 2004

Twelve strains of Bifidobacterium spp. were isolated from the feces of healthy Korean 20∼30 years. The identification of genera from isolates were performed by the microscopic observation and fructose-6-phosphate phosphoketolase (F6PPK) activity which is the key enzyme to distinguish the Bifidobacterium spp. from other anaerobic bacteria. To determine the antibacterial resistance patterns, minimum inhibitory concentration (MIC) of several antibiotics (including anti-tuberculosis agents) was determined. Five of the isolate!, showed the high degree of resistance to vancomycin. To investigate the genetic diversity between isolates and type strain of Bifidobacterium spp. from KCTC, we peformed the RAPD-fingerprinting. Using a total set of four primers, it is possible to distinguish the isolates and Bifidobacterium spp. from KCTC. Thus, Bifidobacterium strains isolated from our samples may be a new species or strains of Bifidobacteriurn genera, and have the potential as a probiotics.

• 한국식품과학회지
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• 제35권5호
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• pp.924-927
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• 2003

비피더스 균주가 가지고 있는 여러 생리 활성을 나타내는 부가가치를 부여한 새로운 고기능성 심치 제품을 개발하기 위해 편성 혐기성 미생물인 비피더스균 가운데 산소에 내성이 있는 Bifidobacterium lactis를 선발하여 김치 제조에 적용하였다. 이 균주의 내산성과 내염성을 각각 알아본 결과 pH 30에서는 별다른 영향을 받지 않았으며 pH 2.5에서도 50% 이상의 생존율을 나타내었다. 그리고 NaCl 농도 3.5%에서도 사멸하지 않는 것으로 나타나 일반적인 김치의 pH와 염농도를 고려할 때 김치에 적용 가능한 것으로 판단되었다. 첨가된 Bifidobacterium lactis가 김치 내에서 생존함을 확인하기 위해서 PCR을 이용하여 확인한 결과 15일 이상 생존하고 있음을 확인할 수 있었다. 그리고 관능평가의 경우 Bifidobacterium lactis 첨가 김치가 전체적인 기호도에서 균주를 첨가하지 않은 대조군 김치와 유사한 특성을 나타냄으로써 Bifidobacterium lactis를 이용한 고기능성 김치의 제조가 가능할 것으로 판단되었다.

• Journal of Microbiology and Biotechnology
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• 제11권5호
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• pp.858-862
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• 2001

The purpose of this research was to investigate the effects of nondigestible oligosaccharides (NDO) from red ginseng marc on the growth of Bifidobacterium spp. Red ginseng marc, a fibrous byproduct of ginseng extract from processing, was destarched by ${\alpha}$-amylase and amyloglucosidase treatment, and then treated with a commercial pectinase to produce NDO. The bifidogenic effects of NDO on B. adolescentis, B. animalis, B. breve, and B. longum were investigated in vitro. NDO significantly promoted the growth of Bifidobacterium spp. The growth, decrease of pH, and organic acid formation (acetate, lactate, formate) were markedly different among the species. B. adolescentis showed the best growth and produced the greatest amount of organic acids. When NDO was used as a carbon source in the cocultivation of Bifidobacterium spp. and Clostridium perfringens, the growth of Bifidobacterium spp. was not influenced by the existence of Cl. perfringens. The result strongly suggested that NDO from red ginseng marc could be used as a potential bifidogenic source.

• Journal of Microbiology and Biotechnology
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• 제17권3호
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• pp.490-495
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• 2007

Lactic acid bacteria (LAB) are beneficial for the gastrointestinal tract and reinforce immunity in human health. Recently, many functional products using the lactic acid bacteria have been developed. Among these LAB, Lactobacillus acidophilus, Lactobacillus rhamnosus, Bifidobacterium longum, and Bifidobacterium bifidum are frequently used for probiotic products. In order to monitor these LAB in commercial probiotic products, a multiplex PCR method was developed. We designed four species-specific primer pairs for multiplex PCR from the 16S rRNA, 16S-23S rRNA intergenic spacer region, and 23S rRNA genes in Lactobacillus acidophilus, Lactobacillus rhamnosus, Bifidobacterium longum, and Bifidobacterium bifidum. Using these primer pairs, 4 different LAB were detected with high specificity in functional foods. We suggest that the multiplex PCR method developed in this study would be an efficient tool for simple, rapid, and reliable identification of LAB used as probiotic strains.

• Journal of Dairy Science and Biotechnology
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• 제22권2호
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• pp.107-112
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• 2004

과산화수소 형태의 반응성 산소에 대한 Lactobacillus Spp., Bifidobacterium spp. 및 Bacillus coagulans의 저항성과 세포 내에서 생성능력을 측정하기 위하여 본 연구가 수행되었다. Lactobacillus spp. 중에서 높은 과산화수소 저항성을 나타낸 균주는 L. acidophilus CU4111와 L. casei Cu4114인 것으로 나타났고 가장 낮은 저항성을 보인 균주는 L. brevis Cu4206이었다. Bifidobacterium longum CU4131은 높은 수준의 저항성을 나타내며 Bacillus coagulans를 포함하는 포자형성유산균주들은 전반적으로 과산화수소에 대한 저항성 이 높은 것으로 나타났다. 세포질 추출액 중의 과산화수소 함유 농도는 Bifidobacterium bifidum CU 4134가 가장 높고 Lactobacilli spp.의 세포질 추출액 중의 과산화수소 함유 농도는 L. casei CU 4114가 가장 높은 것으로 나타났다.

• Food Science and Biotechnology
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• 제16권4호
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• pp.580-583
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• 2007

Phytase activity was examined with various bifidobacterial strains cultured statically in MRS broth at $37^{\circ}C$ for 48 hr. Seven Bifidobacterium species showed mostly an intracellular phytase activity, though their specific activities were very low. The highest specific activity was found in Bifidobacterium animalis B33 strain, among 7 bifidobacteria tested. The specific activity was highest during the exponential growth phase. Carbohydrates and the concentration of phosphorus sources had an effect on the phytase activity and bacterial growth. Glucose was the most favorable carbohydrate for the phytase activity. Phytate inhibited the cell growth, and phytase activity decreased with increase of phytate concentration. The phytase activity was even higher in the static microaerophilic growth than that in anaerobic state, despite the stimulated growth in anaerobic growth. The optimal pH ranges were comparatively broad, but the optimal temperatures were $50^{\circ}C$ for all tested strains. The phytase activity was most active at pH 6.5 and $50^{\circ}C$ for B. animalis B33 strain.