• 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.

• Food Science of Animal Resources
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• v.23 no.3
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• pp.262-268
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• 2003

Kefir is a traditional fermented milk in Caucasusian area and is made mainly of milk fermented with lactic acid bacteria and yeasts. Six typical kefir grains were selected from ten kefir grains collected from different locals in Korea. Kefir grains were gelatinous in texture and had various shapes of villi, grapes, leaves, hulled millets, and towels. To investigate predominant microflora of kefir grains, SPC, MRS, M17, Rogosa, and APT agar media were used for viable cell count MRS, SPC, and Rogosa media were most acceptable for bacterial cell counts of the selected kefir grains. From one or two of the SPC agar plates which contained around 25∼50 colonies, all grown colonies were isolated and identified. Most predominant bacteria was identified as Lactobacillus fermentum by API 50 CHL kit. The proportions of Lb. fermentum and Lb. brevis among the total identified bacteria were around 41~88% and M4%, respectively. To select the best preservation method for kefir grains, refrigeration, freezing, and freeze drying were compared. Freeze drying was found most suitable for the preservation of kefir grains, based upon their acid-producing activities and production of off-flavors.

• Food Science of Animal Resources
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• v.26 no.3
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• pp.402-409
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• 2006

Although microorganisms are, in fact, the most diverse and abundant type of organism on Earth, the ecological functions of microbial populations remains poorly understood. A variety of bacteria including marine Vibrios encounter numerous ecological challenges, such as UV light, predation, competition, and seasonal variations in seawater including pH, salinity, nutrient levels, temperature and so forth. In order to survive and proliferate under variable conditions, they have to develop elaborate means of communication to meet the challenges to which they are exposed. In bacteria, a range of biological functions have recently been found to be regulated by a population density-dependent cell-cell signaling mechanism known as quorum-sensing (QS). In other words, bacterial cells sense population density by monitoring the presence of self-produced extracellular autoinducers (AI). N-acylhomoserine lactone (AHL)-dependent quorum-sensing was first discovered in two luminescent marine bacteria, Vibrio fischeri and Vibrio harveyi. The LuxI/R system of V. fischeriis the paradigm of Gram-negative quorum-sensing systems. At high population density, the accumulated signalstrigger the expression of target genes and thereby initiate a new set of biological activities. Several QS systems have been identified so far. Among them, an AHL-dependent QS system has been found to control biofilm formation in several bacterial species, including Pseudomonas aeruginosa, Aeromonas hydrophila, Burkholderia cepacia, and Serratia liquefaciens. Bacterial biofilm is a structured community of bacterial cells enclosed in a self-produced polymeric matrix that adheres to an inert or living surface. Extracellular signal molecules have been implicated in biofilm formation. Agrobacterium tumefaciens strain NT1(traR, tra::lacZ749) and Chromobacterium violaceum strain CV026 are used as biosensors to detect AHL signals. Quorum sensing in lactic acid bacteria involves peptides that are directly sensed by membrane-located histidine kinases, after which the signal is transmitted to an intracellular regulator. In the nisin autoregulation process in Lactococcus lactis, the NisK protein acts as the sensor for nisin, and NisR protein as the response regulator activatingthe transcription of target genes. For control over growth and survival in bacterial communities, various strategies need to be developed by which receptors of the signal molecules are interfered with or the synthesis and release of the molecules is controlled. However, much is still unknown about the metabolic processes involved in such signal transduction and whether or not various foods and food ingredients may affect communication between spoilage or pathogenic bacteria. In five to ten years, we will be able to discover new signal molecules, some of which may have applications in food preservation to inhibit the growth of pathogens on foods.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.421-431
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• 2016

Recently, poly-γ-glutamic acid synthetase A (pgsA) has been applied to display exogenous proteins on the surface of Lactobacillus casei or Lactococcus lactis, which results in a surface-displayed component of bacteria. However, the ability of carrying genes encoded by plasmids and the expression efficiency of recombinant bacteria can be somewhat affected by the longer gene length of pgsA (1,143 bp); therefore, a truncated gene, pgsA, was generated based on the characteristics of pgsA by computational analysis. Using murine IL-10 as an exogenous gene, recombinant Lactobacillus plantarum was constructed and the capacity of the surface-displayed protein and functional differences between exogenous proteins expressed by these strains were evaluated. Surface expression of IL-10 on both recombinant bacteria with anchorins and the higher expression levels in L. plantarum-pgsA'-IL-10 were confirmed by western blot assay. Most importantly, up-regulation of IL-1β, IL-6, TNF-α, IFN-γ, and the nuclear transcription factor NF-κB p65 in RAW264.7 cells after stimulation with Poly(I:C) or LPS was exacerbated after co-culture with L. plantarum-pgsA. By contrast, IL-10 expressed by these recombinant strains could reduce these factors, and the expression of these factors was associated with recombinant strains that expressed anchorin (especially in L. plantarum-pgsA'-IL-10) and was significantly lower compared with the anchorin-free strains. These findings indicated that exogenous proteins could be successfully displayed on the surface of L. plantarum by pgsA or pgsA', and the expression of recombinant bacteria with pgsA' was superior compared with bacteria with pgsA.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.6
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• pp.787-794
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• 2009

In this study, a new method for extending storage-life and palatability of Betula platyphylla sap by applying lactic acid bacteria fermentation was developed. The fluids of saps were filtered through 0.22 ${\mu}m$ membrane filter and each fermented by 8 different lactic acid bacteria which are Lactobacillus acidophilus, Lactobacillus brevis, Leuconostoc mesenteroides, Leuconostoc lactis, Lactococcus lactis, Pediocossus pentosaceus, Pediococcus dextrinicus, Streptococcus thermophilus. All the tested lactic acid bacteria except P. dextrinicus grew fast up to $10^6{\sim}10^7cfu/mL$ levels and lowered pH down to about pH 4 levels in 48 hours in both saps. The produced organic acids and lowered pH level inhibited the growth of spoilage microorganisms almost completely for 2 weeks during storage at room temperature. Addition of xylitol in the saps before fermentation accelerated the growth of lactic acid bacteria and increased the sweetness and overall taste of final product. The filtration process did not affect the mineral compositions of Betula platyphylla saps. Also the compositions and amounts of minerals showed very minor differences before and after fermentation in Betula platyphylla saps inoculated with L. acidophilus. By applying lactic acid fermentation to extend storage-life of tree saps instead of heat treatment, it was possible to keep natural minerals in active forms without any modifications.

• Journal of Microbiology
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• v.43 no.4
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• pp.345-353
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• 2005

The complex ecosystem of intestinal micro flora is estimated to harbor approximately 400 different microbial species, mostly bacteria. However, studies on bacterial colonization have mostly been based on culturing methods, which only detect a small fraction of the whole microbiotic ecosystem of the gut. To clarify the initial acquisition and subsequent colonization of bacteria in an infant within the few days after birth, phylogenetic analysis was performed using 16S rDNA sequences from the DNA iso-lated from feces on the 1st, 3rd, and 6th day. 16S rDNA libraries were constructed with the amplicons of PCR conditions at 30 cycles and $50^{\circ}C$ annealing temperature. Nine independent libraries were produced by the application of three sets of primers (set A, set B, and set C) combined with three fecal samples for day 1, day 3, and day 6 of life. Approximately 220 clones ($76.7\%$) of all 325 isolated clones were characterized as known species, while other 105 clones ($32.3\%$) were characterized as unknown species. The library clone with set A universal primers amplifying 350 bp displayed increased diversity by days. Thus, set A primers were better suited for this type of molecular ecological analysis. On the first day of the life of the infant, Enterobacter, Lactococcus lactis, Leuconostoc citreum, and Streptococcus mitis were present. The largest taxonomic group was L. lactis. On the third day of the life of the infant, Enterobacter, Enterococcus faecalis, Escherichia coli, S. mitis, and Streptococcus salivarius were present. On the sixth day of the life of the infant, Citrobacter, Clostridium difficile, Enterobacter sp., Enterobacter cloacae, and E. coli were present. The largest taxonomic group was E. coli. These results showed that microbiotic diversity changes very rapidly in the few days after birth, and the acquisition of unculturable bacteria expanded rapidly after the third day.

• Korean Journal of Food Science and Technology
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• v.34 no.3
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• pp.536-539
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• 2002

This study was investigated for the preparation method of meju by three step fermentation. The steamed soybeans were fermented with nisin-producing Lactococcus lactis subsp. lactis IFO12007 to inhibit the growth of odor-producing Bacilli and promote of normal fermentation of meju. The lactie-fermented soybeans were further fermented with Aspergillus oryzae and Bacillus subtilis. The produced meju by this method showed 54%moisture content and pH 7.0, respectively. The three step fermented meju was soaked in 20% (w/v) brine at $25^{\circ}C$ for 90days. After 30 days, the contents of total free amino acids revealed 4,015 mg% which were higher $3{\sim}5$ times than controls. Among the detected free amino acids, the contents of glutamic acid and leucine showed 925 mg% and 380mg%, respectively. Therefore, the new method of producing meju would be an excellent alternative to improve the quality of soybean fermented food, such as doenjang and kanjang.

• Journal of Korean Medicine for Obesity Research
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• v.18 no.2
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• pp.106-114
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• 2018

Objectives: We investigated about the microbial properties and changes in the efficacy of the Codonopsis lanceolata (CL) by natural fermentation. Methods: CL was fermented for four weeks in a well-ventilated place with 2.5% salt. pH, total sugar, total polyphenol, and total flavonoid were measured to determine fermentation characteristics according to fermentation period and salt treatment. Polymerase chain reaction denaturing gradient gel electrophoresis and random amplification of polymorphic DNA-polymerase chain reaction were carried out for microbial analysis during fermentation. In addition, HepG2 cell was cultured to check the lipid accumulation through oil red O staining and the glucose uptake was analyzed by measuring the 2-NBDG at C2C12 cell. Results: The pH level and the total sugar decreased with the CL fermentation. Total polyphenol and flavonoid increased after CL fermentation. It was confirmed that Leuconostoc mesenteroides were maintained continuously during fermentation. In the salt treatment CL, there was a sharp increase in Rahnella aquatilis. Lactobacillus plantarum matrix was observed in fermented CL. In addition, Lactococcus lactis, Weissella koreensis, R. aquatilis, L. plantarum, Leu. mesenteroides have been added to the salt treatment. Glucose uptake were significantly increased after fermentation with salt for four weeks. Lipid accumulation in the HepG2 cells was observed that there was difference (P<0.01) between free fatty acid group (100%) and decreased 4 weeks after fermentation (90.38%) at $800{\mu}g/mL$. Conclusions: Total polyphenol and flavonoid were increased after CL fermentation. Especially, percentage of the glucose uptake and lipid accumulation inhibition increased in CL fermentation with salt. It is expected that fermentation of salt treated CL will be more effective in diabetes and fatty liver.

• Journal of Convergence for Information Technology
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• v.11 no.4
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• pp.111-121
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• 2021

In order to determine the possibility that Chrysanthemum indicum L. cultured with Lactococcus lactis (CILL) is a material for obesity suppression food, the body weight, body fat mass, and T cells were determined in C57BL/6 mice induced by a high fat diet. The CILL (25.15±2.44 g) demonstrated weight loss from week 4 onward and maintained a low weight gain from week 1 to week 8 (1.00±0.53 g). The 8-week body weight (30.38±4.17 g) indicated loss of 13.15% when compared to the HFD (60% high fat diet, 34.99±2.09 g). Fat mass decreased to 10.3022±2.8813 g, and the absolute liver weight decreased relative to that in the HFD. CD4+ T cells were 4.84±1.33%, CD8+ T cells 7.02±2.26%, and CD4+CD8+ T cells 1.46±0.81%, which were all higher than those in the HFD. As a result, CILL can be used as a material for preventing obesity as an effective measure toward reducing weight when consumed orally.

• Animal Bioscience
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• v.35 no.2
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• pp.281-289
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• 2022

Objective: The aim of this study was to characterize the exopolysaccharides (EPS)-producing lactic acid bacteria from Taiwanese ropy fermented milk (TRFM) for developing a clean label low-fat fermented milk. Methods: Potential isolates from TRFM were selected based on the Gram staining test and observation of turbid suspension in the culture broth. Random amplified polymorphic DNA-polymerase chain reaction, 16S rRNA gene sequencing, and API CHL 50 test were used for strain identification. After evaluation of EPS concentration, target strains were introduced to low-fat milk fermentation for 24 h. Fermentation characters were checked: pH value, acidity, viable count, syneresis, and viscosity. Sensory evaluation of fermented products was carried out by 30 volunteers, while the storage test was performed for 21 days at 4℃. Results: Two EPS-producing strains (APL15 and APL16) were isolated from TRFM and identified as Lactococcus (Lc.) lactis subsp. cremoris. Their EPS concentrations in glucose and lactose media were higher than other published strains of Lc. lactis subsp. cremoris. Low-fat fermented milk separately prepared with APL15 and APL16 reached pH 4.3 and acidity 0.8% with a viable count of 9 log colony-forming units/mL. The physical properties of both products were superior to the control yogurt, showing significant improvements in syneresis and viscosity (p<0.05). Our low-fat products had appropriate sensory scores in appearance and texture according to sensory evaluation. Although decreasing viable cells of strains during the 21-day storage test, low-fat fermented milk made by APL15 exhibited stable physicochemical properties, including pH value, acidity, syneresis and sufficient viable cells throughout the storage period. Conclusion: This study demonstrated that Lc. lactis subsp. cremoris APL15 isolated from TRFM had good fermentation abilities to produce low-fat fermented milk. These data indicate that EPS-producing lactic acid bacteria have great potential to act as natural food stabilizers for low-fat fermented milk.