• Journal of Animal Science and Technology
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• v.65 no.3
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• pp.652-663
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• 2023

The rumen fluids contain a wide range of bacteria, protozoa, fungi, and viruses. The various ruminal microorganisms in the rumen provide nutrients by fermenting the forage they eat. During metabolic processes, microorganisms present in the rumen release diverse vesicles during the fermentation process. Therefore, in this study, we confirmed the function of rumen extracellular vesicles (EVs) and their interaction with the host. We confirmed the structure of the rumen EVs by transmission electron microscope (TEM) and the size of the particles using nanoparticle tracking analysis (NTA). Rumen EVs range in size from 100 nm to 400 nm and are composed of microvesicles, microparticles, and ectosomes. Using the Caenorhabditis elegans smart animal model, we verified the interaction between the host and rumen EVs. Exposure of C. elegans to rumen EVs did not significantly enhance longevity, whereas exposure to the pathogenic bacteria Escherichia coli O157:H7 and Staphylococcus aureus significantly increased lifespan. Furthermore, transcriptome analysis showed gene expression alterations in C. elegans exposed to rumen EVs, with significant changes in the metabolic pathway, fatty acid degradation, and biosynthesis of cofactors. Our study describes the effect of rumen EV interactions with the host and provides novel insights for discovering biotherapeutic agents in the animal industry.

• Food Science of Animal Resources
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• v.44 no.1
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• pp.146-164
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• 2024

Owing to the residual toxicity and adverse health effects of chemical preservatives, there is an increasing demand for using natural preservatives in food. Although many natural extracts have been evaluated, research on their antibacterial effects remains insufficient. Therefore, this study aimed to explore the possibility of developing Psidium guajava, Ecklonia cava, and Paeonia japonica (Makino) Miyabe & Takeda extracts as natural food preservatives. Further, the effect of mixing these extracts on microbial growth and quality was evaluated during the refrigeration of sausages. Optimal mixing ratios were determined based on the minimum inhibitory and bactericidal concentrations of each mixed extract against the Listeria monocytogenes, Salmonella spp. and Escherichia coli. D-optimal mixing design optimization tool was further used to obtain an optimum mixing ratio of Formulation 1 (F1). The antibacterial activity of F1 increased with increasing concentration, with similar activities at 0.5% and 1%. The sausages with synthetic or natural preservatives showed significantly lower lipid oxidation than those of the control and grapefruit extract-treated sausages after 4 wk of refrigeration. Total plate counts were observed only in the control and treatment groups stored for 3 wk, and no significant effect of ascorbic acid was observed. Compared to the other samples, sausages with added natural extracts showed the highest overall acceptability scores initially and after 4 wk. Therefore, similar amounts of grapefruit seed and natural extracts had the same effect on microbiological analysis and lipid rancidity during sausage storage. Hence, this mixture can serve as a potential natural preservative in meat products.

• Journal of Microbiology and Biotechnology
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• v.33 no.11
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• pp.1506-1512
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• 2023

Quantitative analysis of adenosine triphosphate (ATP) has been widely used as a diagnostic tool in the food and medical industries. Particularly, the pathogenesis of a few diseases including inflammatory bowel disease (IBD) is closely related to high ATP concentrations. A bioluminescent D-luciferin/luciferase system, which includes a luciferase (FLuc) from the firefly Photinus pyralis as a key component, is the most commonly used method for the detection and quantification of ATP. Here, instead of isolating FLuc produced in recombinant Escherichia coli, we aimed to develop a whole-cell biocatalyst system that does not require extraction and purification of FLuc. To this end, the gene coding for FLuc was introduced into the genome of probiotic Saccharomyces boulardii using the CRISPR/Cas9-based genome editing system. The linear relationship (r² = 0.9561) between ATP levels and bioluminescence generated from the engineered S. boulardii expressing FLuc was observed in vitro. To explore the feasibility of using the engineered S. boulardii expressing FLuc as a whole-cell biosensor to detect inflammation biomarker (i.e., ATP) in the gut, a colitis mouse model was established using dextran sodium sulfate as a colitogenic compound. Our findings demonstrated that the whole-cell biosensor can detect elevated ATP levels during gut inflammation in mice. Therefore, the simple and powerful method developed herein could be applied for non-invasive IBD diagnosis.

• Journal of Animal Science and Technology
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• v.66 no.1
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• pp.103-114
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• 2024

This study was done to investigate the effects of the incorporation of Achyranthes japonica extracts (AJE) in diet on the production parameters of growing pigs. Exp 1: Total, 105 crossbred pigs (average body weight: 24.47 ± 2.46 kg) were used in a 6-week feeding trial. Pigs (seven replicates, five pigs per pen) were allotted randomly to three treatments. Dietary treatments: CON (basal diet); basal diet with 0.025% AJE, and basal diet + 0.050% AJE). Growth performance, nutrient digestibility, fecal microbial count, and fecal noxious gas were assessed in this study. Average daily gain (ADG), average daily feed intake (ADFI), and gain to feed ratio (G:F) were not affected by the addition of up to 0.05% AJE. In the case of apparent total tract digestibility (ATTD), dry matter (DM), nitrogen (N), and digestible energy (DE) were not changed in 3rd and 6th weeks of the feeding trial through the addition of AJE up to 0.05% in the growing pig diet. In microbial count, Lactobacillus and Escherichia coli count at 3rd and 6th week was similar in all the treatment diets. The inclusion of AJE at levels up to 0.05% in growing pig diet had no effect on the production of NH3, H2S, acetic acid, and CO₂ in the feces. After ending the Exp 1, a total of nine pigs were divided into three treatment groups. Treatment diets were included, TRT1, basal diet + powder quercetin 30 g; TRT2, basal diet + powder quercetin 150 g; TRT3, basal diet + powder quercetin 300g. Rate of absorption in blood was increased with the higher dose of quercetin. The results suggested incorporation of AJE up to 0.05% has no significant effect on ADG, ADFI, and G:F, as well as DM, N, and DE digestibility, fecal microbial count, and fecal noxious gas emission in growing pigs, even though no negative effect was found.

• IMMUNE NETWORK
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• v.24 no.3
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• pp.27.1-27.14
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• 2024

The tumor microenvironment (TME) is formed by several immune cells. Notably, tumor-associated macrophages (TAMs) are existed in the TME that induce angiogenesis, metastasis, and proliferation of cancer cells. Recently, a point-mutated variant of IL-32θ was discovered in breast cancer tissues, which suppressed migration and proliferation through intracellular pathways. Although the relationship between cancer and IL-32 has been previously studied, the effects of IL-32θ on TAMs remain elusive. Recombinant human IL-32θ (rhIL-32θ) was generated using an Escherichia coli expression system. To induce M0 macrophage polarization, THP-1 cells were stimulated with PMA. After PMA treatment, the cells were cultured with IL-4 and IL-13, or rhIL-32θ. The mRNA level of M1 macrophage markers (IL-1β, TNFα, inducible nitric oxide synthase) were increased by rhIL-32θ in M0 macrophages. On the other hand, the M2 macrophage markers (CCL17, CCL22, TGFβ, CD206) were decreased by rhIL-32θ in M2 macrophages. rhIL-32θ induced nuclear translocation of the NF-κB via regulation of the MAPK (p38) pathway. In conclusion, point-mutated rhIL-32θ induced the polarization to M1-like macrophages through the MAPK (p38) and NF-κB (p65/p50) pathways.

• Journal of Life Science
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• v.29 no.6
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• pp.671-678
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• 2019

Antibacterial and antioxidant activities of plant sources have attracted a wide range of interest across the world over the last decade. This is due to the growing concern for safe and alternative sources of antibacterial and antioxidant agents. In this study, we focused on the antibacterial and antioxidant activities and the chemical composition of a methanol extract from Viburnum sargentii seeds. The chemical composition was determined by gas chromatography-mass spectroscopy (GC-MS), and the antibacterial activity was screened by a disc diffusion assay. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined using the microbroth dilution and spread plate method, respectively. The V. sargentii extract showed growth inhibition activity on all tested Gram-positive (Listeria monocytogenes, Staphylococcus aureus, and Staphylococcus saprophyticus) and Gram-negative (Escherichia coli, Pseudomonas putida, and Proteus vulgaris) pathogenic bacteria. The MIC and MBC ranged from 0.156~1.25 mg/ml for Gram-positive and 0.625~5.0 mg/ml for Gram-negative tested bacteria. The GC-MS results revealed the presence of several phytochemicals such as ${\beta}-sitosterol$ and vitamin E, which are known for their pharmacological applications. The antioxidant activities of V. sargentii extract were investigated by three different methods: the 2,2-diphenyl-1-picrylhydrazyl free radical scavenging assay, the reducing power assay, and the total antioxidant capacity assay. The results showed a concentration-dependent antioxidant potential for all three used methods. In sum, our findings suggest that the methanol extract of V. sargentii seeds has the potential to inhibit the growth of pathogenic bacteria and provide antioxidant compounds, making it therefore worthy of further investigation.

• Korean Journal of Food Science and Technology
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• v.38 no.5
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• pp.699-706
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• 2006

The antioxidative and antimicrobial activities of Euphorbia jolkini extracts were investigated. Total polyphenohc compounds extracted were approximately as follows: 162.08 mg/g from ethanol, 12.64 mg/g from n-hexane, 48.11 mg/g from dichloromethane, 544.08 mg/g from ethyl acetate, 176.42 mg/g from butanol, and 30.00 mg/g from water. The ethylacetate fraction of this extraction showed the highest antioxidative activity $(IC_{50})$: DPPH radical scavenging capacity was measured at $8.38\;{\mu}g/mL$, xanthine oxidase inhibitory activity was $466.01\;{\mu}g/mL$, superoxide radical scavenging capacity was $11.39\;{\mu}g/mL$, and nitric oxide scavenging capacity was $332.11\;{\mu}g/mL$. Antimicrobial activities were determined by paper disc method and minimum inhibitory concentration of E. jolkini extracts against food-borne pathogens and spoilage bacteria. The growth inhibition curves of E. jolkini extracts against Bacillus cereus, Listeria monocytogenes, and Escherichia coli were also determined. These results suggest that the ethylacetate fraction of E. jolkini has strong antimicrobial activity against the all species of microorganisms as well as strong antioxidant activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.9
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• pp.1106-1112
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• 2007

The solvent extracts of Euphorbia helioscopia, which were extracted by using several solvents with different polarities, were prepared for utility as natural preservatives. The E. helioscopia extract by 80% ethanol was sequentially fractionated with n-hexane, dichloromethane, ethylacetate, and butanol. In order to effectively screen for a natural preservatives agent, we first investigated the antioxidant activities such as DPPH radical scavenging capacity, superoxide radical scavenging capacity, and xanthine oxidase inhibitory activity of the E. helioscopia extracts. By the screening system, we found that ethylacetate fraction had the strongest antioxidant activity in a dose-dependent manner. The antimicrobial activities and cell growth inhibition were investigated for each strain with the different concentrations of E. helioscopia extracts. Antimicrobial activities were shown in ethylacetate fraction of E. helioscopia; however, ethanol, butanol and water fractions showed weak antimicrobial activity against the tested microorganisms. Among the five fractions, ethylacetate fraction showed the highest antimicrobial activities against microorganisms tested, such as Bacillus sublitis, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella Enteritidis and Salmonella Typhimurium. The polyphenol content from ethanol, n-hexane, dichloromethane, ethylacetate, butanol, and water fractions were 207.46 mg/g, 45.45 mg/g, 138.23 mg/g, 678.02 mg/g, 278.91 mg/g, and 63.76 mg/g, respectively. There seems to be a close relationship between antioxidant activities, and antimicrobial activities and polyphenol content in natural plant. From these results, it is suggested that E. helioscopia could be used for the ethylacetate fraction and could be suitable for the development of a food preservative.

• Journal of Life Science
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• v.16 no.3 s.76
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• pp.485-492
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• 2006

The PrrBA two-component system is one of the major regulatory systems that control expression of photosynthesis genes in response to changes in oxygen tension in the anoxygenic photosynthetic bacterium, Rhodobacter sphaeroides. The system consists of the PrrB histidine kinase and the PrrA response regulator. The N-terminal transmembrane domain of PrrB serves as a signal-sensing domain and comprises six transmembrane helices forming three periplasmic loops and two cytoplasmic loops. The $3^{rd}$ and $4^{th}$ transmembrane helices and the $2^{nd}$ periplasmic loop were suggested to play a crucial role in redox-sensory function. In this study we demonstrated that mutations of Asp-90, Gln-93, Leu-94, Leu-98, and Asn-106 in the $2^{nd}$ periplasmic loop and its neighboring region led to severe defects in PrrB sensory function, indicating that these amino acids might be related to the redox-sensing function of PrrB. The mutant forms (D90E, D90N, and D90A) of PrrB were heterologously overexpressed in Escherichia coli, purified by means of affinity chromatography and their autokinase activities were comparatively assessed. The D90N form of PrrB was shown to possess higher autokinase activity than the wild-type form of PrrB, whereas the D90E form of PrrB displayed lower autokinase activity than the wild-type form of PrrB. The D90A mutation led to the loss of PrrB autokinase activity.

• Korean Journal of Microbiology
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• v.47 no.4
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• pp.308-315
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• 2011

The purpose of this work was to examine the antimicrobial activity derived from the lactic acid bacterium, UK-3 isolated from the vaginas of women of childbearing age. Various physiological and biochemical properties of this strain were characterized. Both the BIOLOG system and phylogenetic analysis using 16S rRNA sequencing were utilized for identification, and the strain was designated as Lactobacillus plantarum UK-3, and registered in GenBank as [JK266589]. Growth rate, production of organic acids (e.g., lactic acid and acetic acid), and pH during growth were monitored. The maximum concentrations of lactic acid and acetic acid were approximately 684.11 mM and 174.26 mM, respectively, and pH changed from 7.0 to 3.7 after 72 h of incubation. High performance liquid chromatography was used to confirm lactic acid and acetic acid production. Significant antimicrobial activity of the concentrated supernatant was demonstrated against various Gram-positive (e.g., Staphylococcus aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus, Enterococcus faecalis, Neisseria species., Listeria monocytogenes), Gram-negative bacteria (e.g., Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis), and yeast (e.g., Candida albicans) by the plate diffusion method. As a result, the concentrated L. plantarum UK-3 cultures had lower acidity and inhibited the growth of all microorganisms tested, whereas the growth of L. acidophilus was not affected.