• Food Science and Biotechnology
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• v.15 no.4
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• pp.559-563
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• 2006

The growth responses of six bacterial strains exposed to materials extracted from turmeric (Curcuma longa) rhizomes were examined using impregnated paper disk agar diffusion. Methanol extracts of turmeric rhizomes exhibited strong inhibitory activity against Clostridium perfringens and weak inhibitory activity toward Escherichia coli at 5 mg/disk. However, in tests conducted with Bifidobacterium adolescentis, B. bifidum, B. longum, and Lactobacillus casei, the methanol extract showed no inhibitory response. The biologically active constituent isolated from the turmeric rhizomes extracts was characterized as ar-turmerone using various spectroscopic analyses including EI-MS and NMR. The responses varied according to the dosage, chemicals, and bacterial strain tested. At 2 and 1 mg/disk, ar-turmerone strongly inhibited the growth of C. perfringens and moderately inhibited the growth of E. coli without any adverse effects on the growth of four lactic acid-bacteria. Of the commercially available compounds originating from turmeric rhizomes, curcumin exhibited strong and moderate growth inhibition against C. perfringens at 2 and 1 mg/disk, respectively, and weak growth inhibition against E. coli at 1 mg/disk. However, little or no activity was observed for borneol, 1,8-cineole, and sabinene against all six bacteria strains tested. The observed inhibitory activity of the turmeric rhizome-derived curcumin and ar-turmerone against C. perfringens and E. coli demonstrate one of the important pharmacological activities of turmeric rhizomes.

• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.610-616
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• 2002

The growth-inhibiting effects of Pinus densiflpora leaf-derived materials on nine human intestinal bacteria were investigated using the impregnated paper disk method, and their activities were compared with those of 13 commercially available terpenes. The biologically active constituent of the extract of P densiflora leaf was characterized as the monoterpene (1R)-(+)-$\alpha$-pinene by various spectroscopic analyses. Responses varied according to bacterial strain, chemicals, and dose. At 10 mg/disk, limonene and (1R)-(+)-$\alpha$-pinene strongly inhibited the growth of Clostridium perfringens, Staphylococcus aureus, and Escherichia coli, without adverse effects on the growth of five lactic acid-bacteria (Bifidobacterium adolescentis, B. bifidum, B. longum, Lactobacillus acidophilus, and L. casei). Little or no inhibition against seven bacteria was observed with anethole, borneol, camphor, caryophyllene, 1,8-cineole, estragole, linalool, and $\alpha$-terpineol. Structure-activity relationship revealed that (1R)-(+)-$\alpha$-pinene had more growth-inhibiting activity against C. perfringens than (1R)-(+)-$\beta$-, (1S-(-)-$\alpha$-, and (1S-(-)-$\beta$-pinenes. Furthermore, the growth-inhibition against L. casei was much more pronounced in (1R)-(+)-$\beta$- and (In-(-)-$\beta$-pinenes than (1R)-(+)-$\alpha$- and (1S)-(-)-$\alpha$-pinenes. These results indicate that the (+)-$\alpha$ form seems to be required against C. perfringens and $\beta$ form against L. casei for growth-inhibiting activity. Morphologically, most strains of C. perfringens were damaged and disappeared at 5 and 2 mg/disk of (1R)-(+)-$\alpha$-pinene. Morphological study revealed that (1R)-(+)-$\alpha$-pinene had more growth-inhibiting activity against C. perfringens than (1R)-(+)-$\beta$-, (1S)-(-)-$\alpha$-, and (1S)-(-)-$\beta$-pinenes. As naturally occurring growth-inhibiting agents, the Pinus leaf-derived materials described above could be useful preventive agents against diseases caused by harmful intestinal bacteria such as clostridia.

• Journal of Ginseng Research
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• v.14 no.2
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• pp.253-264
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• 1990

The growth responses of a variety of human intestinal bacteria to extracts of Panax ginseng and five other oriental medicinal Araliaceae were elraluattd in vitro and in vivo. The extracts enhanced the growth of Brifidobnnerilrm breve and B. longlim in media with or without carbon sources, suggesting that bifidus factors) might be involved in the phenomenon. This effect was most pronounced with water extract of P. ginseng, the growth of 27 bifidobacteria strains belonging to B adolescentis, B. longum, B. brim and B. infantis being greatly stimurated, whereas seven B. bifidum strains and other bacteria such as clostridia and Escherichin soli had little or no ability to utilise it for growth. Methanol extracts of p. ginseng were found to selectively inhibit growth of various clostridia including bifidobacteria. Paraputrificum, but this effect was not observed on other bacteria including bifidobacteria. The effect of ginseng extract intake (600 mg/day for two weeks) on the faecal microflora, pH, volatile fatty acids, ammonia, putrefactive products, and -glucuronidase, -glucosidase and nitroreductase activities, and on the blood components (triglyceride, total cholesterol and ammonia) were investigated using seven healthy human volunteers. The total concentration of faecal microflora including Bifidnkaderiifm app. during the period of ginseng extract intake %twas significantly unaffected from the preceding and subsequent control peroids. However, the frequency of occurrence of subjects having C. perfringens was significantly decreased. The faecal pH value was also significantly decreased, suggesting that the intake might increase the activity of Bifidobncterium spry. Other biochemical properties in faeces did not changed significantly. The levels of ammonia and triglycerid in blood were decreased with ginseng extract intake. These results may be an indication of at least one of the Pharmacological actions of p. ginseng as an adaptogen.

• Journal of Microbiology and Biotechnology
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• v.13 no.4
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• pp.529-536
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• 2003

The growth-inhibitory activity of Cassia tora seed-derived materials against seven intestinal bacteria was examined in vitro, and compared with that of anthraquinone, anthraflavine, anthrarufin, and 1-hydroxyanthraquinone. The active constituent of C. tore seeds was characterized as quinizarin, using various spectroscopic analyses. The growth responses varied depending on the compound, dose, and bacterial strain tested. At 1 mg/disk, quinizarin exhibited a strong inhibition of Clostridium perfringens and moderate inhibition of Staphylococcus aureus without any adverse effects on the growth of Bifidobacterium adolescentis, B. bifidum, B. longum, and Lactobacillus casei. Furthermore, the isolate at 0.1 mg/disk showed moderate and no activity against C. perfringens and S. aureus. The structure-activity relationship revealed that anthrarufin, anthraflavine, and quinizarin moderately inhibited the growth of S. aureus. However. anthraquinone and 1-hydroxyanthraquinone did not inhibit the human intestinal bacteria tested. As for the morphological effect of 1 mg/disk quinizarin, most strains of C. perfringens were damaged and disappeared, indicating that the strong activity of quinizarin was morphologically exhibited against C. perfringens. The inhibitory effect on aflatoxin $B_1$ biotransformation by anthraquinones revealed that anthrarufin ($IC_50,\;11.49\mu\textrm{M}$) anthraflavine ($IC_50,\;26.94\mu\textrm{M}$), and quinizarin ($IC_50,\;4.12\mu\textrm{M}$), were potent inhibitors of aflatoxin ${B_1}-8,9-epoxide$ formation. However, anthraquinone and 1-hydroxyanthraquinone did not inhibit the mouse liver microsomal sample to convert aflatoxin $B_1$ to aflatoxin ${B_1}-8,9-epoxide$. These results indicate that the two hydroxyl groups on A ring of anthraquinones may be essential for inhibiting the formation of aflatoxin ${B_1}-8,9-epoxide$. Accordingly, as naturally occurring inhibitory agents, the C. tora seed-derived materials described could be useful as a preventive agent against diseases caused by harmful intestinal bacteria, such as clostridia, and as an inhibitory agent for the mouse liver microsomal conversion of aflatoxin $B_1$ to aflatoxin ${B_1}-8,9-epoxide$.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.491-497
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• 1992

The water extract of Sinomeniiacuti Radix (Sinomenium acutum) was found out to have a strong inhibition activity on the growth of Clostridium perfringens. The anti-bacterial activity was stable at the range of pH 1 to pH 13 and kept in a thermal treatment at $121^{\circ}C$ for 15 minutes. The minmal inhibition concentration of the Sinomeniiacuti Radix extract on the growth of Cl. peifringens was 1000 ppm. The Sinomeniiacuti Radix extract also suppressed the growth of Cl. ramosum, Cl. paraputnficum, Cl. butyn'cum, Bifidobacterium blficum, Bacteriodes fragilis, Eubacterium limosum. The extract, however, did not inhibit the growth of Bif adolescentis, Blf. infantis, Bif longum, E. coli. Enterococcus faecalis and Staphylococcus aureus. On the other hand, the extract showed a promoting effect for the growth of Bif animalis and Lactobacillus acidophilus.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.535-540
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• 2012

${\beta}$-D-Xylosidase (E.C. 3.2.1.37) from Bifidobacterium breve K-110, which hydrolyzes ginsenoside Ra1 to ginsenoside Rb2, was cloned and expressed in Escherichia coli. The ($His_6$)-tagged recombinant enzyme, designated as XlyBK-110, was efficiently purified using $Ni^{2+}$-affinity chromatography (109.9-fold, 84% yield). The molecular mass of XylBK-100 was found to be 55.7 kDa by SDS-PAGE. Its sequence revealed a 1,347 bp open reading frame (ORF) encoding a protein containing 448 amino acids, which showed 82% identity (DNA) to the previously reported glycosyl hydrolase family 30 of Bifidobacterium adolescentis ATCC 15703. The $K_m$ and $V_{max}$ values toward p-nitrophenyl-${\beta}$-D-xylopyranoside (pNPX) were 1.45mM and 10.75 ${\mu}mol/min/mg$, respectively. This enzyme had pH and temperature optima at 6.0 and $45^{\circ}C$, respectively. XylBK-110 acted to the greatest extent on xyloglucosyl kakkalide, followed by pNPX and ginsenoside Ra1, but did not act on p-nitrophenyl-${\alpha}$-L-arabinofuranoside, p-nitrophenyl-${\beta}$-D-glucopyranoside, or p-nitrophenyl-${\beta}$-D-fucopyranoside. In conclusion, this is the first report on the cloning and expression of ${\beta}$-D-xylosidase-hydrolyzing ginsenoside Ra1 and kakkalide from human intestinal microflora.

• Journal of Applied Biological Chemistry
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• v.42 no.4
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• pp.202-204
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• 1999

Methanol extracts of leaves from 15 Pinus species belonging to the family Pinaceae were tested for their in vitro growth-inhibiting activities against 10 bacteria commonly found in the gastrointestinal tracts of human, using impregnated paper disk methods. The inhibitory activities varied with both bacterial strain and Pinus species used. At a concentration of 10 mg/disk, a clear growth inhibition was produced from the extracts of Pinus armandii, P. banksiana, P. bungeana, P. densiflora, P. rigida, and P. thunbergii against Clostridium perfringens, whereas all Pinus samples revealed weak or little growth-inhibiting activity against Escherichia coli, Bacteroides fragilis, and Staphylococcus aureus. At 5 mg/disk, the extracts of P. banksiana and P. thunbergii exhibited potent growth inhibition toward C. perfringens. All the extracts except the one from P. densiflora did not adversely affect growth of Bifidobacterium adolescentis, B. longum, B. bifidum, B. breve, B. animalis, and Lactobacillus casei. The growth-inhibiting activity was more pronounced in C. perfringens, as compared to the lactic acid-producing bacteria. These results may be an indication of at least one of the pharmacological activities of these Pinus species.

• Food Science and Biotechnology
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• v.16 no.1
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• pp.99-103
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• 2007

The purpose of this study was to develop a simple detection method, possibly at the species-level, that allows for large-scale screening of bifidobacteria. Human fecal samples were plated on MRS-raffinose agar containing cysteine and neomycin sulfate, serving as selective pressure for bifidobacteria, and 0.003%(w/v) bromocresol green. All of the test strains grew well on this medium at $37{\pm}1^{\circ}C$, forming white colonies surrounded by yellow halos, which presented a sharp contrast against the green background. In this disc assay, the required incubation time to develop a yellowish zone varied with the species of Bifidobacterium that was tested, allowing for differential counts and easy identification at the species-level: 10-14 hr for B. bifidum, 20-22 hr for B. catenulatum and B. infantis. and 24-25 hr for B. longum and B. breve. No apparent color was observed for B. angulatum and B. adolescentis 28 hr after inoculation. To evaluate the results of pH indicator-based identification, individual isolates were subjected to a colony-PCR experiment with genus-specific primers. The amplified products from the isolates were in good accordance with those from the reference strains at a level of 95% agreement. These results suggest that the present method could be conveniently applied to cell counts, as well as to the preliminary identification of bifidobacteria from a variety of sample types including human feces, dairy products, and commercial probiotic supplements.

• Korean Journal of Microbiology
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• v.49 no.3
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• pp.245-252
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• 2013

Pseudomonas aeruginosa and Acinetobacter baumannii are significant opportunistic pathogens in hospitals and are resistant to most antibiotics. Multidrug-resistant P. aeruginosa (MDRPA) and A. baumannii (MDRAB) cause severe human nosocomial infections and are more difficult to treat than methicillin-resistant Staphylococcus aureus (MRSA). Bifidobacteria are among of the most beneficial probiotics and have been widely studied for their antimicrobial activities. The present study explored the antimicrobial activity of Bifidobacterium sp. isolated from healthy Koreans against MDRPA and MDRAB. The antimicrobial activity of the isolates against MDRPA and MDRAB, which are resistant to ciprofloxacin, tobramycin, gentamicin, meropenem, and ceftazidime, was determined by modified broth microdilution methods using absorbance. Among all tested bifidobacteria isolates (nine B. adolescentis, three B. longum, and two B. pseudocatenulatum), the culture supernatant of B. pseudocatenulatum SPM1309 showed a strong growth inhibitory effect against MDRPA and MDRAB. No change in the turbidity of the mixture was observed during incubation, and its inhibitory effect occurred through bacteriostastic action. Moreover, the antibacterial activity was observed in the fraction with molecular weights <10 kDa of bifidobacteria culture supernatant, and the active fraction was heat-stable because it maintained its activity when heated at $70^{\circ}C$ for 10 min. The results suggest that this Bifidobacterium strain could have potential applications for alternative therapy in MDRPA and MDRAB infections.

• Food Science of Animal Resources
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• v.40 no.6
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• pp.1014-1032
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• 2020

This work aimed to investigate the psychobiotic effects of six bacterial strains on the mind and behavior of male Wistar rats. The probiotic (PRO) group (n=7) were rats pre-treated with antibiotics for 7 days followed by 14-day probiotic administration, antibiotics (ANT) group (n=7) were rats treated with antibiotics for 21 days without probiotics. The control (CON) group (n=7) were rats that received sham treatment for 21 days. The six bacterial strains with probiotic properties were mostly isolated from Thai fermented foods; Pedicoccus pentosaceus WS11, Lactobacillus plantarum SK321, L. fermentum SK324, L. brevis TRBC 3003, Bifidobacterium adolescentis TBRC 7154 and Lactococcus lactis subsp. lactis TBRC 375. The probiotics were freeze-dried into powder (6×10⁹ CFU/5 g) and administered to the PRO group via oral gavage. Behavioral tests were performed. The PRO group displayed significantly reduced anxiety level and increased locomotor function using a marble burying test and open field test, respectively and significantly improved short-term memory performance using a novel object recognition test. Antibiotics significantly reduced microbial counts in rat feces in the ANT group by 100 fold compared to the PRO group. Probiotics significantly enhanced antioxidant enzymatic and non-enzymatic defenses in rat brains as assessed using catalase activity and ferric reducing antioxidant power assay, respectively. Probiotics also showed neuroprotective effects with less pyknotic cells and lower frequency of vacuolization in cerebral cortex. This multi-strain probiotic formulation from Thai fermented foods may offer a potential to develop psychobiotic-rich functional foods to modulate human mind and behaviors.