• Title/Summary/Keyword: CFU assay

Search Result 176, Processing Time 0.025 seconds

Protective Efficacy of Recombinant Proteins Adenylate Kinase, Nucleoside Diphosphate Kinase, and Heat-Shock Protein 70 against Mycobacterium tuberculosis Infection in Mice (유전자 재조합 단백질 Adenylate Kinase, Nucleoside Diphosphate Kinase와 Heat-Shock Protein 70의 결핵균에 대한 방어면역효능 분석)

  • Lee, Seung-Heon;Lee, Eun-Gae;Kim, Su-Yeon;Cho, Sang-Nae;Park, Young-Kil;Bai, Gill-Han
    • Tuberculosis and Respiratory Diseases
    • /
    • v.58 no.2
    • /
    • pp.142-152
    • /
    • 2005
  • Background : Priming and boosting vaccination strategy has been widely explored for new vaccine development against tuberculosis. As an effort to identify other vaccine candidates, this study was initiated to evaluate protective efficacy of adenylate kinase (AK), nucleoside diphosphate kinase (NdK), and heat shock protein 70 (Hsp70) of Mycobacterium tuberculosis. Method : M. tuberculosis genes encoding AK, NdK, and Hsp70 proteins were amplified by PCR and cloned into E. coli expression vector, pQE30. Recombinant AK, NdK, and Hsp70 was purified through Ni-NTA resin. To evaluate immune responses, we performed enzyme-linked immunosorbent assay (ELISA) for IgG isotype and $IFN-{\gamma}$ after mice were immunized subcutaneously with recombinant proteins delivered in dimethyl dioctadecylammonium bromide (DDA). Immunized- and control groups were challenged by aerosol with M. tuberculosis. The spleens and lungs of mice were removed aseptically and cultured for CFU of M. tuberculosis. Result : Vaccination with recombinant proteins AK, NdK, and Hsp70 delivered in DDA elicited significant level of antibody and $IFN-{\gamma}$ responses to corresponding antigens but no protective immunity comparable to that achieved with Mycobacterium bovis BCG. Conclusion : Recombinant proteins AK, NdK, and Hsp70 do not effectively control growth of M. tuberculosis in mice when immunized with DDA as an adjuvant.

Effects of Dietary Probiotics Supplementation on Juvenile Olive Flounder Paralichthys olivaceus (치어기 넙치 사료내 생균제 첨가효과)

  • Jeong Chang-Wha;Choi Hee-Jung;Yoo Gwang-Yeol;Lee Seung-Hyung;Kim Young-Chul;OKorie Okorie Eme;Lee Jun-Ho;Jun Kyoung-Dong;Choi Se-Min;Kim Kang-Woong;Kang Yong-Jin;Kang Ju-Chan;Kong In-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.39 no.6
    • /
    • pp.460-465
    • /
    • 2006
  • An 8-week feeding trial was conducted to investigate the effects of dietary supplementation with probiotics as a feed additive for Juvenile olive flounder (Paralichthys olivaceus). Three experimental diets supplemented with Bacillus polyfermenticus (BP), Bacillus licheniformis (BL), or Bacillus polyfermenticus plus Saccharomyces cerevisiae, (BP+SC) at $1.0{\times}10^7CFU/kg$ diet on a dry-matter basis were prepared. The basal diet was used as a control. After the 8-week feeding trial, the respiratory burst activity (NBT assay) of fish fed the BP + SC diet was significantly higher than that of fish fed the control diet. Fish fed the BP, BL and BP + SC diets had significantly lower cumulative mortality than did fish fed the control diet after the third day of the challenge test (P<0.05). However, there were no significant differences among fish fed the experimental diets in weight gain, feed efficiency, protein efficiency ratio, hematosomatic index, condition factor, survival rate, or Iysozyme activity. Results could suggest that dietary B. polyfermenticus, B. licheniformis, and B. polyfermenticus +S. cerevisiae enhance nonspecific immunity and disease resistance in juvenile olive flounder.

Effects of propolis extracts on the immune response in cultured flounder, Paralichthys olivaceus (Propolis extracts가 양식넙치의 면역활성능에 미치는 영향)

  • Nam, Hyun Ju;Park, Kyung Il;Choi, Min Soon
    • Journal of fish pathology
    • /
    • v.27 no.1
    • /
    • pp.47-56
    • /
    • 2014
  • Propolis is a beehive product with a very complex chemical composition, widely used in folk medicine because of its several therapeutic activities. This study was conducted to measure the efficacy of propolis on non-specific defense reactions, specific immune response, and protection levels against pathogen challenge with Streptococcus iniae. in vitro and in vivo. In vitro, the phagocytic activity and NBT assay of peripheral blood leukocytes (PBL) were evaluated in a various propolis extractsconcentrations (0, 10, 50, 100, 150, 250 and $500{\mu}g/ml$). The optimal concentration showing activation of propolis extracts was determined to $100{\mu}g/ml$. In vivo, they were divided into four groups (PBS, propoli extractss, vaccine, propolis extracts + vaccine) in vivo. Fish were received i.p. injection of either PBS or propolis extracts, and in the presence or absence of formalin inactivated S. iniae ($1{\times}10^8$ CFU/fish), respectively. The level of haematocrit is not affected among experimental groups. The phagocytic activity and the NBT reduction activities of head kidney phagocyte were markedly (p<0.05)augmented in the propolis extracts groups than in the PBS-control group, respectively. The level of serum lysozyme activity was significantly (p<0.05) increased in the propolis extracts treated groups than in the PBS-control group. The agglutinin titer was significantly (p<0.05) enhanced in the vaccine+propolis extracts group than in the vaccine group, but there was no difference between PBS-control and propolis treated group. The results of the present study suggest that propolis extracts seems to be a promising compounds of non-specific immune stimulator, also being able to use a good adjuvant.

Agronomic characteristics and field resistance to bacterial soft rot of transgenic potato overexpressing the soybean calmodulin 4 gene (SCaM4) (SCaM4 과발현 형질전환 감자의 농업적 특성 및 감자 무름병 저항성 평가)

  • Sohn, Hwangbae;Cho, Kwangsoo;Cho, Jihong;Gwon, Ohgeun;Cheon, Chunggi;Choi, Jigyeong;Chung, Woosik;Lee, Shin Woo
    • Journal of Plant Biotechnology
    • /
    • v.39 no.4
    • /
    • pp.295-299
    • /
    • 2012
  • We performed in vitro assay and field trials to assess levels of changes in intrinsic properties and resistance against soft rot of the potato cv. Dejima upon the introduction of a soybean calmodulin 4 gene (SCaM4). Field trials with four lines overexpressing SCaM4 gene were conducted over two seasons, and harvested tubers were evaluated in bioassay for resistance to Pectobacterium carotovorum ssp. carotovorum. The SCaM4 transgenic potato lines inoculated with $10^8$ CFU/ml of P. carotovorum ssp. carotovorum showed enhanced resistance compared to control. Among the SCaM4 transgenic lines, the transgenic line SCaM4-4 exhibited the highest tolerance to soft rot in vitro assays, so did in field trials. In the field trial, the soft rot resistance of SCaM4-4 line was more than 5 times higher compared to that of control cultivar, Dejima. The major agronomic characteristics of the SCaM4 transgenic lines were not different from those of the nontransgenic 'Dejima'. The result demonstrated that the transformation of a calmodulin 4 gene was a successful strategy in development of potato cultivar enhanced to soft rot.

Psychobiotic Effects of Multi-Strain Probiotics Originated from Thai Fermented Foods in a Rat Model

  • Luang-In, Vijitra;Katisart, Teeraporn;Konsue, Ampa;Nudmamud-Thanoi, Sutisa;Narbad, Arjan;Saengha, Worachot;Wangkahart, Eakapol;Pumriw, Supaporn;Samappito, Wannee;Ma, Nyuk Ling
    • Food Science of Animal Resources
    • /
    • v.40 no.6
    • /
    • pp.1014-1032
    • /
    • 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.

Effect of Reduction in the Adipose Accumulation of Akkermansia muciniphila in Mature 3T3-L1 Adipocytes (성숙한 3T3-L1 지방세포에서 Akkermansia muciniphila의 지방축적 감소 효과)

  • Shim, Hyeyoon;Lim, Sookyoung;Shin, Joo-Hyun;Lee, Dokyung;Seo, Jae-Gu;Kim, Hojun
    • Journal of Korean Medicine for Obesity Research
    • /
    • v.19 no.2
    • /
    • pp.106-112
    • /
    • 2019
  • Objectives: The aim of this study was to observe the reduction of lipid accumulation by treatment with Akkermansia muciniphila extract on 3T3-L1 adipocytes. Methods: After treating pasteurized Akk. muciniphila strains in HT-29 colorectal cancer cell, the relative expression of interleukin (IL)-8, tumor necrosis factor-α, IL-6, and IL-1β mRNA was analyzed by real time polymerase chain reaction, respectively. 27 strains of Akk. muciniphila which have anti-inflammatory effects were selected. 3T3-L1 pre-adipocytes were treated with Akk. muciniphila for 24 hr and then measured the toxicity using water soluble tetrazolium salt assay. The cells were incubated for 4 days and then differentiated into adipocytes using the medium including adipogenic reagents for 10 days. The Akk. muciniphila was treated when the medium was exchanged for differentiation medium at 4th day and insulin medium at 6th day. To observe the lipid accumulation, the cells were stained with Oil red O dye and were measured using a spectrophotometer. Results: In the cytotoxicity test, the cell viability of 3T3-L1 pre-adipocytes was significantly increased compared to the control group which untreated with Akk. muciniphila, and there was no cytotoxicity of Akk. muciniphila at 1×107 CFU/mL. The results on Oil red O staining and absorbance measurements were showed a significant decrease in lipid accumulation in the group which was treated with Akk. muciniphila compared to the control group. Conclusions: In our results, Akk. muciniphila has the inhibitory effect of lipid accumulation in 3T3-L1 adipocytes. This suggests that Akk. muciniphila could be help to improve obesity.

Antimicrobial Effects of Ethanol Extract of Yongdamgosam-hwan against Streptococcus mutans (용담고삼환(龍膽苦參丸) 에탄올 추출물의 Streptococcus mutans에 대한 항균활성에 관한 연구)

  • Yun, Yong-Il;Lee, Hae-Soo;Jung, Min-Ji;You, Seong-Il;Song, Yung-Sun;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
    • /
    • v.30 no.6
    • /
    • pp.55-61
    • /
    • 2015
  • Objectives : Yongdamgosam-hwan(YGH) has been used as a traditional medicine from old times for antiinflammatory effects. Streptococcus mutans(S. mutans) is known as a prime bacteria responsible for causing caries by forming a biofilm referred to as dental plaque on the tooth surface. But antimicrobial activity of YGH with dental disease is not sufficiently understood. This study was designed to investigate the effects of YGH ethanol extract on antimicrobial effect against Streptococcus mutans.Methods : The antimicrobial effect of YGH ethanol extract was assessed by the paper disk diffusion method and optical density method to determine minimum inhibition concentration(MIC), also observed by fractional inhibitory concentration index(FICI) and time-kill assay to figure out the synergic effect on the combination of YGH ethanol extract with antibiotics.Results : The YGH ethanol extract 500 μg was 7.5-8.5 mm diameter of clear zone of inhibition against Streptococcus mutans in a concentration-dependent manner and MIC was 250 μg/mL. The administration of the ethanol extract in combination with gentamicin and streptomycin induced a reduction of ≥4-8-fold in all tested bacteria. Furthermore, time-kill study was found that a combination of YGH ethanol extract with oxacillin and streptomycin produced a more rapid decrease in the concentration of bacteria CFU/mL than the YGH ethanol extract or antibiotics alone.Conclusions : As a result, the YGH ethanol extract has good antimicrobial effects. And the results suggest that YGH could be employed as a natural antibacterial agent in dental care products.

Probiotic Properties of Lactic Acid Bacteria isolated from Feces and Kimchi (베트남인 분변 및 김치로부터 분리된 유산균의 프로바이오틱스 기능성 연구)

  • Shin, Hyun Su;Yoo, Sung Ho;Jang, Jin Ah;Won, Ji Young;Kim, Cheol Hyun
    • Journal of Dairy Science and Biotechnology
    • /
    • v.35 no.4
    • /
    • pp.255-261
    • /
    • 2017
  • The purpose of this study was to investigate the probiotic properties and antioxidant capacity of lactic acid bacteria isolated from Vietnamese feces and the Korean traditional food kimchi. Six isolated strains were identified as Lactobacillus sp. by 16S rRNA sequencing. All strains showed good resistance to low pH (1.5, 2.0, and 3.0) and 0.3% oxgall bile acids. Culture filtrates from the six strains showed various antioxidant effects, including DPPH, ABTS, reducing power, and metal chelating ($Fe^{2+}$) activities. Two of the six Lactobacillus strains showed potential probiotic activity. Heat resistance and adhesion assays were conducted by mixing the selected strains, Lactobacillus acidophilus V4, Lactobacillus plantarum V7, and Lactobacillus paracasei DK121 isolated from kimchi. The results showed that the heat resistance of these strains was similar to that of a commercial strain, L. plantarum LP. In addition, a mucin attachment assay using the mixture of selected strains (V4, V7, and DK121) showed high binding activity to the mucous layer. In conclusion, a mixture of V4, V7, and DK121 shows promising probiotic activity and may be useful for the development of health-related products.

Antibacterial effect of electrolyzed water on Streptococcus mutans (전기분해수소수의 Streptococcus mutans에 대한 항균효과)

  • Kim, Ji-Hye;Youn, Ha-Young;Kim, Eun-Kyong;Lee, Young-Eun;Jang, Ji-Eon;Song, Keun-Bae
    • Journal of Korean society of Dental Hygiene
    • /
    • v.21 no.5
    • /
    • pp.527-533
    • /
    • 2021
  • Objectives: Electrolyzed water has been proven to have antibacterial effects against various microorganisms. However, there are only a few studies about effects of electrolyzed water on oral bacteria. The purpose of this study was to examine the antibacterial effect of electrolyzed water on Streptococcus mutans in vitro. Methods: S. mutans KCOM 1054 was treated with electrolyzed water for 1 or 3 minutes and plated on Mitis Salivarius agar with 15% sucrose and bacitracin. After incubation for 48 hours, colony forming units (CFU) were counted, and dental plaque was quantified by crystal violet staining. Results: The growth of S. mutans was significantly inhibited by electrolyzed water (p<0.001). In addition, the dental plaque formation by S. mutans was decreased in a time-dependent manner by exposure to electrolyzed water (p<0.001). Conclusions: Our results suggest that electrolyzed water can effectively prevent dental caries by inhibiting growth of (and the formation of dental plaque by) S. mutans.

Preventive effects of shiitake mushroom extract on candida stomatitis (칸디다성 구내염에 대한 표고버섯 추출물의 예방효과)

  • Yoo, Hyun-Jun
    • Journal of Dental Rehabilitation and Applied Science
    • /
    • v.37 no.3
    • /
    • pp.123-129
    • /
    • 2021
  • Purpose: The purpose of this study was to investigate antifungal activity of shiitake mushroom yeast and hyphal type of Candida albicans. Materials and Methods: The extract from shiitake mushroom was collected by drying the supernatant after soaking shiitake mushrooms in water or ethanol. The antifungal activity of the extracts against yeast type of C. albicans was investigated by the susceptibility assay using microplate. C. albicans biofilm was formed on 12-well plate using Ham's F-12 medium in CO2 incubator and treated with the ethanol extract. Furthermore, C. albicans biofilm was formed on denture base resin disk and treated with or without the ethanol extract in the presence of denture cleanser. Live C. albicans in biofilm was counted by cultured colony forming unit value after inoculated on agar plate. Results: Ethanol extract from shiitake mushroom showed stronger antifungal activity against yeast type of C. albicans compared to its water extract. The ethanol extract significantly reduced count of C. albicans in hyphal biofilm (P < 0.05). Also, the ethanol extract showed synergistically antifungal effect with denture cleanser on candidal biofilm on denture base resin disk (P < 0.05). Conclusion: The ethanol extract of shiitake mushroom may be a candidate for preventing candidal stomatitis as well as denture-related stomatitis.