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Showing papers in "Journal of Microbiology and Biotechnology in 2019"


Journal ArticleDOI
TL;DR: This review focused on the relationship between probiotics and the human gut microbiota and its roles in gut microbiome-associated diseases and discussed future directions and research areas that need further elucidation in order to better understand the roles of probiotics in the treatment of intestinal diseases.
Abstract: Probiotics, including bacteria and yeast, are live microorganisms that have demonstrated beneficial effects on human health. Recently, probiotic bacteria are constantly being studied and their applications are also being considered in promising adjuvant treatments for various intestinal diseases. Clinical trials and in vivo experiments have extended our current understanding of the important roles that probiotics play in human gut microbiomeassociated diseases. It has been documented through many clinical trials that probiotics could shape the intestinal microbiota leading to potential control of multiple bowel diseases and promotion of overall wellness. In this review, we focused on the relationship between probiotics and the human gut microbiota and its roles in gut microbiome-associated diseases. Here, we also discuss future directions and research areas that need further elucidation in order to better understand the roles of probiotics in the treatment of intestinal diseases.

165 citations


Journal ArticleDOI
TL;DR: Age-related rearrangement of gut microbiota was observed in centenarians, such as reduced proportions of Faecalibacterium and Prevotella, and increased proportion of Escherichia, along with higher abundances of Akkermansia, Clostridium, Collinsella, and uncultured Christensenellaceae.
Abstract: Several studies have attempted to identify factors associated with longevity and maintenance of health in centenarians. In this study, we analyzed and compared the gut microbiota of centenarians in longevity villages with the elderly and adults in the same region and urbanized towns. Fecal samples were collected from centenarians, elderly, and young adults in longevity villages, and the gut microbiota sequences of elderly and young adults in urbanized towns of Korea were obtained from public databases. The relative abundance of Firmicutes was found to be considerably higher in subjects from longevity villages than those from urbanized towns, whereas Bacteroidetes was lower. Age-related rearrangement of gut microbiota was observed in centenarians, such as reduced proportions of Faecalibacterium and Prevotella, and increased proportion of Escherichia, along with higher abundances of Akkermansia, Clostridium, Collinsella, and uncultured Christensenellaceae. Gut microbiota of centenarians in rehabilitation hospital were also different to those residing at home. These differences could be due to differences in diet patterns and living environments. In addition, phosphatidylinositol signaling system, glycosphingolipid biosynthesis, and various types of N-glycan biosynthesis were predicted to be higher in the gut microbiota of centenarians (corrected p < 0.05). These three metabolic pathways of gut microbiota can be associated with the immune status and healthy gut environment of centenarians. Although further studies are necessary to validate the function of microbiota between groups, this study provides valuable information on centenarians' gut microbiota.

70 citations


Journal ArticleDOI
TL;DR: Recent progress in the development of synthetic biology tools for the production of novel secondary metabolites in Streptomyces is reviewed, including genomic elements and genome engineering tools for StrePTomyces, the heterologous gene expression strategy of designed biosynthetic gene clusters in the Streptomeces chassis strain, and future directions to expand diversity of novelsecondary metabolites.
Abstract: Streptomyces are attractive microbial cell factories that have industrial capability to produce a wide array of bioactive secondary metabolites. However, the genetic potential of the Streptomyces species has not been fully utilized because most of their secondary metabolite biosynthetic gene clusters (SM-BGCs) are silent under laboratory culture conditions. In an effort to activate SM-BGCs encoded in Streptomyces genomes, synthetic biology has emerged as a robust strategy to understand, design, and engineer the biosynthetic capability of Streptomyces secondary metabolites. In this regard, diverse synthetic biology tools have been developed for Streptomyces species with technical advances in DNA synthesis, sequencing, and editing. Here, we review recent progress in the development of synthetic biology tools for the production of novel secondary metabolites in Streptomyces, including genomic elements and genome engineering tools for Streptomyces, the heterologous gene expression strategy of designed biosynthetic gene clusters in the Streptomyces chassis strain, and future directions to expand diversity of novel secondary metabolites.

55 citations


Journal ArticleDOI
TL;DR: Analysis of recent studies on the assembly systems of OM components show that OM assembly machines have the potential to be a novel attractive drug target of Gram-negative bacteria.
Abstract: Gram-negative pathogens, such as Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii, pose a serious threat to public health worldwide, due to high rates of antibiotic resistance and the lack of development of novel antimicrobial agents targeting Gram-negative bacteria. The outer membrane (OM) of Gram-negative bacteria is a unique architecture that acts as a potent permeability barrier against toxic molecules, such as antibiotics. The OM is composed of phospholipids, lipopolysaccharide (LPS), outer membrane β-barrel proteins (OMP), and lipoproteins. These components are synthesized in the cytoplasm or in the inner membrane, and are then selectively transported to the OM by the specific transport machines, including the Lol, BAM, and Lpt pathways. In this review, we summarize recent studies on the assembly systems of OM components and analyze studies for the development of inhibitors that target these systems. These analyses show that OM assembly machines have the potential to be a novel attractive drug target of Gram-negative bacteria.

54 citations


Journal ArticleDOI
TL;DR: The results indicated that JW15 possesses anti-inflammatory potential and provide a molecular basis regarding the development of functional probiotic products.
Abstract: Probiotics are known to provide the host with immune-modulatory effects and are therefore of remarkable interest for therapeutic and prophylactic applications against various disorders, including inflammatory diseases. Weissella cibaria JW15 (JW15) has been reported to possess probiotic and antioxidant properties. However, the effect of JW15 on inflammatory responses has not yet been reported. Therefore, the objective of the current study was to evaluate the anti-inflammatory potential of JW15 against lipopolysaccharide (LPS) stimulation. The production of pro-inflammatory factors and the cellular signaling pathways following treatment with heat-killed JW15 was examined in LPS-induced RAW 264.7 cells. Treatment with heat-killed JW15 decreased nitric oxide and prostaglandin E2 production via downregulation of the inducible nitric oxide synthase and cyclooxygenase-2. In addition, treatment with heat-killed JW15 suppressed the expression of pro-inflammatory cytokines, interleukin (IL)-1β, IL-6, and tumor necrosis factor-α. The anti-inflammatory properties of treating with heat-killed JW15 were associated with mitogen-activated protein kinase signaling pathwaymediated suppression of nuclear factor-κB. These results indicated that JW15 possesses antiinflammatory potential and provide a molecular basis regarding the development of functional probiotic products.

53 citations


Journal ArticleDOI
TL;DR: Ahead of Print article withdrawn by publisher; further details to be confirmed.
Abstract: UCB-1 is the commercial rootstock of pistachio. Reproduction of this rootstock by tissue culture is limited by low levels of proliferation rate. Therefore, any compound that improves the proliferation rate and the quality of the shoots can be used in the process of commercial reproduction of this rootstock. Use of plant growth-promoting bacteria is one of the best ideas. Given the beneficial effects of nanoparticles in enhancement of the growth in plant tissue cultures, the aim of the present study was to investigate the effects of nanoencapsulation of plant growth-promoting rhizobacteria (using silica nanoparticles and carbon nanotubes) and their metabolites in improving UCB1 pistachio micropropagation. The experiment was conducted in a completely randomized design with three replications. Before planting, treatments on the DKW medium were added. The results showed that the use of Pseudomonas fluorescens VUPF5 and Bacillus subtilis VRU1 nanocapsules significantly enhanced the root length and proliferation. The nanoformulation of the VUPF5 metabolite led to the highest root length (6.26 cm) and the largest shoot (3.34 cm). Inoculation of explants with the formulation of the metabolites (both bacterial strains) significantly elevated the average shoot length and the fresh weight of plant compared to the control. The explants were dried completely using both bacterial strains directly and with capsule coating after the three days.

43 citations


Journal ArticleDOI
YunJung Lee1, Yu Ra Ji1, Sumi Lee, Mi-Jung Choi1, Youngjae Cho1 
TL;DR: In this paper, a probiotic Lactobacillus acidophilus KBL409 was encapsulated with alginate and Alginate-chitosan (Al/Chi) through extrusion method.
Abstract: The probiotic Lactobacillus acidophilus KBL409 was encapsulated with alginate (Al) and alginate-chitosan (Al/Chi) through extrusion method. The sizes and zeta potentials of microspheres were measured to confirm encapsulation. To evaluate the protective effect of microspheres against gastrointestinal fluids, all the samples were exposed to simulated gastric fluids (SGFs, pH 1.5) at 37°C for 1 or 2 h, followed by incubation with simulated intestinal fluids (SIFs, pH 6.5) for 2 h. The mucoadhesive ability of microspheres was evaluated using the intestinal epithelial cell line HT29-MTX. To extend the shelf-life of probiotics, lyoprotectants such as disaccharide and polysaccharide were mixed with free or encapsulated cells during the freeze-drying process. The size of the microspheres demonstrated a narrow distribution, while the zeta potentials of Al and Al/Chi-microspheres were -17.9 ± 2.3 and 20.4 ± 2.6 mV, respectively. Among all the samples, Al/Chi-encapsulated cells showed the highest survival rate even after exposure to SGF and SIF. The mucoadhesive abilities of Al and Al/Chi-microspheres were higher than 94%, whereas the free L. acidophilus showed 88.1% mucoadhesion. Ten percent of sucrose showed over 80% survival rate in free or encapsulated cells. Therefore, L. acidophilus encapsulated with Al and Al/Chi-microspheres showed higher survival rates after exposure to the gastrointestinal tract and better mucoadhesive abilities than the free cells. Also, sucrose showed the highest protective effect of L. acidophilus during the freeze-drying process.

39 citations


Journal ArticleDOI
TL;DR: It is noteworthy that ChNP can be also applied to deliver antimicrobial drug, which further enhance the efficiency and stability of the antimicrobial agent.
Abstract: Polymeric nanoparticles are widely used for drug delivery due to their biodegradability property. Among the wide array of polymers, chitosan has received growing interest among researchers. It was widely used as a vehicle in polymeric nanoparticles for drug targeting. This review explored the current research on the antimicrobial activity of chitosan nanoparticles (ChNP) and the impact on the clinical applications. The antimicrobial activities of ChNP were widely reported against bacteria, fungi, yeasts and algae, in both in vivo and in vitro studies. For pharmaceutical applications, ChNP were used as antimicrobial coating for promoting wound healing, preventing infections and combating the rise of infectious disease. Besides, ChNP also exhibited significant inhibitory on foodborne microorganisms, particularly on fruits and vegetables. It is noteworthy that ChNP can be also applied to deliver antimicrobial drugs, which further enhance the efficiency and stability of the antimicrobial agent. The present review addresses the potential antimicrobial applications of ChNP from these few aspects.

37 citations


Journal ArticleDOI
TL;DR: It is demonstrated that strains H19-1 and H20-5 induced ABA-independent and dependent salinity tolerance, respectively, in tomato plants, therefore these strains can be used as effective bio-fertilizers for sustainable agriculture.
Abstract: Salinity is one of the major abiotic stresses that cause reduction of plant growth and crop productivity. It has been reported that plant growth-promoting bacteria (PGPB) could confer abiotic stress tolerance to plants. In a previous study, we screened bacterial strains capable of enhancing plant health under abiotic stresses and identified these strains based on 16s rRNA sequencing analysis. In this study, we investigated the effects of two selected strains, Bacillus aryabhattai H19-1 and B. mesonae H20-5, on responses of tomato plants against salinity stress. As a result, they alleviated decrease in plant growth and chlorophyll content; only strain H19-1 increased carotenoid content compared to that in untreated plants under salinity stress. Strains H19-1 and H20-5 significantly decreased electrolyte leakage, whereas they increased Ca2+ content compared to that in the untreated control. Our results also indicated that H20-5-treated plants accumulated significantly higher levels of proline, abscisic acid (ABA), and antioxidant enzyme activities compared to untreated and H19-1-treated plants during salinity stress. Moreover, strain H20-5 upregulated 9-cisepoxycarotenoid dioxygenase 1 (NCED1) and abscisic acid-response element-binding proteins 1 (AREB1) genes, otherwise strain H19-1 downregulated AREB1 in tomato plants after the salinity challenge. These findings demonstrated that strains H19-1 and H20-5 induced ABA-independent and -dependent salinity tolerance, respectively, in tomato plants, therefore these strains can be used as effective bio-fertilizers for sustainable agriculture.

34 citations


Journal ArticleDOI
TL;DR: It is reported that ORF4a, ORf4b, and ORF8b proteins, alone or in combination, effectively antagonize nuclear factor kappa B (NF-κB) activation and are involved in viral escape of NF-κBs-mediated antiviral immune responses.
Abstract: Middle East respiratory syndrome coronavirus (MERS-CoV) is a newly emerging coronavirus which is zoonotic from bats and camels. Its infection in humans can be fatal especially in patients with preexisting conditions due to smoking and chronic obstructive pulmonary disease (COPD). Among the 25 proteins encoded by MERS-CoV, 5 accessory proteins seem to be involved in viral evasion of the host immune responses. Here we report that ORF4a, ORF4b, and ORF8b proteins, alone or in combination, effectively antagonize nuclear factor kappa B (NF-κB) activation. Interestingly, the inhibition of NF-κB by MERS-CoV accessory proteins was mostly at the level of pattern recognition receptors: melanoma differentiationassociated gene 5 (MDA5). ORF4a and ORF4b additively inhibit MDA5-mediated activation of NF-κB while that of retinoic acid-inducible gene 1 (RIG-I) is largely not perturbed. Of note, ORF8b was found to be a novel antagonist of MDA5-mediated NF-kκB activation. In addition, ORF8b also strongly inhibits Tank-binding kinase 1 (TBK1)-mediated induction of NF-κB signaling. Taken together, MERS-CoV accessory proteins are involved in viral escape of NF-κB-mediated antiviral immune responses.

33 citations


Journal ArticleDOI
TL;DR: It is suggested that NK41 and NK46 may alleviate anxiety/depression and colitis by suppressing gut dysbiosis by suppressing Gut Proteobacteria and Bacteroidetes populations and bacterial lipopolysaccharide production.
Abstract: We isolated Lactobacillus mucosae NK41 and Bifidobacterium longum NK46 from human feces, which induced BDNF expression in corticosterone-stimulated SH-SY5Y cells, and examined their anti-depressive effects in mice. NK41, NK46, and their (1:1) mixture significantly mitigated immobilization stress (IS)-induced anxiety-like/depressive behaviors, hippocampal NF-κB activation, BDNF expression, Iba1+ cell population, and blood corticosterone, TNF-α, IL- 6, and lipopolysaccharide levels. Furthermore, they inhibited colitis marker NF-κB activation, and TNF-α expression in mice with IS-induced anxiety/depression. They additionally suppressed gut Proteobacteria and Bacteroidetes populations and bacterial lipopolysaccharide production. These findings suggest that NK41 and NK46 may alleviate anxiety/depression and colitis by suppressing gut dysbiosis.

Journal ArticleDOI
TL;DR: Puffing may be an effective and economical technique for enhancing the extraction yield and levels of minor ginsenosides responsible for the major biological activities of ginseng.
Abstract: Korean ginseng (Panax ginseng Meyer) was processed by drying, steaming, or puffing, and the effects of these processes on the ginsenoside profile were investigated. The main root of 4-year-old raw Korean ginseng was dried to produce white ginseng. Steaming, followed by drying, was employed to produce red or black ginseng. In addition, these three varieties of processed ginseng were puffed using a rotational puffing gun. Puffed ginseng showed significantly higher extraction yields of ginsenosides (49.87-58.60 g solid extract/100 g of sample) and crude saponin content (59.40-63.87 mg saponin/g of dried ginseng) than nonpuffed ginseng, respectively. Moreover, puffing effectively transformed the major ginsenosides (Rb1, Rb2, Rc, Rd, Re, and Rg1) of ginseng into minor ones (F2, Rg3, Rk1, and Rg5), comparable to the steaming process effect on the levels of the transformed ginsenosides. However, steaming takes much longer (4 to 36 days) than puffing (less than 30 min) for ginsenoside transformation. Consequently, puffing may be an effective and economical technique for enhancing the extraction yield and levels of minor ginsenosides responsible for the major biological activities of ginseng.

Journal ArticleDOI
Wei Tang1, Yue Wang1, Zhang Jun1, Yali Cai1, Zengguo He1 
TL;DR: The practical approaches to improving carotenoid yields, which have been facilitated by advancements in strain work as well as the optimization of media and fermentation conditions, were summarized respectively.
Abstract: Rhodotorula is a group of pigment-producing yeasts well known for its intracellular biosynthesis of carotenoids such as β-carotene, γ-carotene, torulene and torularhodin. The great potential of carotenoids in applications in food and feed as well as in health products and cosmetics has generated a market value expected to reach over $2.0 billion by 2022. Due to growing public concern over food safety, the demand for natural carotenoids is rising, and this trend significantly encourages the use of microbial fermentation for natural carotenoid production. This review covers the biological properties of carotenoids and the most recent findings on the carotenoid biosynthetic pathway, as well as strategies for the metabolic engineering methods for the enhancement of carotenoid production by Rhodotorula. The practical approaches to improving carotenoid yields, which have been facilitated by advancements in strain work as well as the optimization of media and fermentation conditions, were summarized respectively.

Journal ArticleDOI
TL;DR: Considering its high fibrinolytic activity, significant salt tolerance, and ability to grow at 10°C, B. velezensis BS2 can be used as a starter for jeotgal.
Abstract: Bacillus sp. BS2 showing strong fibrinolytic activity was isolated from sea squirt (munggae) jeotgal, a traditional Korean fermented seafood. BS2 was identified as B. velezensis by molecular biological methods.B. velezensis BS2 grows well at 15% NaCl and at 10oC. When B. velezensis BS2 was cultivated in TSB broth for 96 h at 37°C, the culture showed the highest fibrinolytic activity (131.15 mU/µl) at 96 h. Three bands of 27, 35 and 60 kDa were observed from culture supernatant by SDS-PAGE, and fibrin zymography showed that the major fibrinolytic protein was the 27 kDa band. The gene (aprEBS2) encoding the major fibrinolytic protein was cloned, and overexpressed in heterologous hosts, B. subtilis WB600 and E. coli BL21 (DE3). B. subtilis transformant showed 1.5-fold higher fibrinolytic activity than B. velezensis BS2. Overproduced AprEBS2 in E. coli was purified by affinity chromatography. The optimum pH and temperature were pH 8.0 and 37°C, respectively. Km and Vmax were 0.15 mM and 39.68 µM/l/min, respectively, when N-succinyl-Ala-Ala-Pro-Phe-pNA was used as the substrate. AprEBS2 has strong α-fibrinogenase and moderate β-fibrinogenase activity. Considering its high fibrinolytic activity, significant salt tolerance, and ability to grow at 10°C, B. velezensis BS2 can be used as a starter for jeotgal.

Journal ArticleDOI
TL;DR: The aim of this review is to summarize potential strategies applicable for mitigating AR at the environment-livestock interface.
Abstract: The rise of antimicrobial resistance (AR) is a major threat to global health. The food animal industry contributes to the increasing occurrence of AR. Multiple factors can affect the occurrence and dissemination of AR in the animal industry, including antibiotic use and farm management. Many studies have focused on how the use of antibiotics in food-producing animals has led to the development of AR. However, a few effective mitigating strategies for AR have been developed in food-producing animals, especially those exposed to the environment. The aim of this review is to summarize potential strategies applicable for mitigating AR at the environment-livestock interface.

Journal ArticleDOI
TL;DR: In a semi-continuous cultivation system, a hydraulic retention time of 5 days with sludge inoculation resulted in the highest biomass productivity and N/P removal, by introducing bacteria in activated sludge at optimized timing.
Abstract: Although chemical oxygen demand (COD) is an important issue for wastewater treatment, COD reduction with microalgae has been less studied compared to nitrogen or phosphorus removal. COD removal is not efficient in conventional wastewater treatment using microalgae, because the algae release organic compounds, thereby finally increasing the COD level. This study focused on enhancing COD removal and meeting the effluent standard for discharge by optimizing sludge inoculation timing, which was an important factor in forming a desirable algae/bacteria consortium for more efficient COD removal and higher biomass productivity. Activated sludge has been added to reduce COD in many studies, but its inoculation was done at the start of cultivation. However, when the sludge was added after 3 days of cultivation, at which point the COD concentration started to increase again, the algal growth and biomass productivity were higher than those of the initial sludge inoculation and control (without sludge). Algal and bacterial cell numbers measured by qPCR were also higher with sludge inoculation at 3 days later. In a semi-continuous cultivation system, a hydraulic retention time of 5 days with sludge inoculation resulted in the highest biomass productivity and N/P removal. This study achieved a further improved COD removal than the conventional microalgal wastewater treatment, by introducing bacteria in activated sludge at optimized timing.

Journal ArticleDOI
TL;DR: Findings reveal that endophytic fungi from the rubber tree F. elastica leaves exhibit unique characteristics and are potential producers of novel natural bioactive products.
Abstract: Endophytic fungi are an important component of plant microbiota, and have the excellent capacity for producing a broad variety of bioactive metabolites. These bioactive metabolites not only affect the survival of the host plant, but also provide valuable lead compounds for novel drug discovery. In this study, forty-two endophytic filamentous fungi were isolated from Ficus elastica leaves, and further identified as seven individual taxa by ITS-rDNA sequencing. The antimicrobial activity of these endophytic fungi was evaluated against five pathogenic microorganisms. Two strains, Fes1711 (Penicillium funiculosum) and Fes1712 (Trichoderma harzianum), displayed broad-spectrum bioactivities. Our following study emphasizes the isolation, identification and bioactivity testing of chemical metabolites produced by T. harzianum Fes1712. Two new isocoumarin derivatives (1 and 2), together with three known compounds (3-5) were isolated, and their structures were elucidated using NMR and MS. Compounds 1 and 2 exhibited inhibitory activity against Escherichia coli. Our findings reveal that endophytic fungi from the rubber tree F. elastica leaves exhibit unique characteristics and are potential producers of novel natural bioactive products.

Journal ArticleDOI
TL;DR: This study reported for the first time the effectiveness of P. chlororaphis to control BLSB of maize and revealed that KNU17Pc1 had strong in vitro antifungal activity and was effective toward all in vitro PGP traits except phosphate solubilization.
Abstract: In this study, strain KNU17Pc1 was tested for its antifungal activity against Rhizoctonia solani AG-1(IA), which causes banded leaf and sheath blight (BLSB) of maize. KNU17Pc1 was tested further for its broad-spectrum antifungal activity and in vitro plant growth promoting (PGP) traits. In addition, the in vivo effects of KNU17Pc1 on reduction of BLSB severity and seedling growth promotion of two maize cultivars under greenhouse conditions were investigated. On the basis of multilocus sequence analysis (MLSA), KNU17Pc1 was confirmed as P. chlororaphis subsp. aurantiaca. The study revealed that KNU17Pc1 had strong in vitro antifungal activity and was effective toward all in vitro PGP traits except phosphate solubilization. In this study, for the first time, a strain of P. chlororaphis against Colletotrichum dematium, Colletotrichum gloeosporioides, Fusarium oxysporum f.sp. melonis, Fusarium subglutinans and Stemphylium lycopersici has been reported. Further biochemical studies showed that KNU17Pc1 was able to produce both types of phenazine derivatives, PCA and 2-OH-PCA. In addition, solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) analysis identified 13 volatile organic compounds (VOCs) in the TSB culture of KNU17Pc1, 1-undecene being the most abundant volatile. Moreover, for the first time, Octamethylcyclotetrasiloxan (D4), dimethyl disulfide, 2-methyl-1,3-butadiene and 1-undecene were detected in P. chlororaphis. Furthermore, this study reported for the first time the effectiveness of P. chlororaphis to control BLSB of maize. Hence, further studies are necessary to test the effectiveness of KNU17Pc1 under different environmental conditions so that it can be exploited further for biocontrol and plant growth promotion.

Journal ArticleDOI
TL;DR: The composition of the fecal microbiome in dogs is affected by breed, and the relative abundance of Firmicutes was significantly lower in the Maltese dog breed than that in the other two breeds, while that of Fusobacteria was significantly higher in the maltese than in the Miniature Schnauzer breed.
Abstract: The gut microbiome influences the health and well-being of dogs. However, little is known about the impact of breed on the fecal microbiome composition in dogs. Therefore, we aimed to investigate the differences in the fecal microbiome in three breeds of dog fed and housed under the same conditions, namely eight Maltese (8.0 ± 0.1 years), eight Miniature Schnauzer (8.0 ± 0.0 years), and nine Poodle dogs (8.0 ± 0.0 years). Fresh fecal samples were collected from the dogs and used to extract metagenomic DNA. The composition of the fecal microbiome was evaluated by 16S rRNA gene amplicon sequencing on the MiSeq platform. A total of 840,501 sequences were obtained from the 25 fecal samples and classified as Firmicutes (32.3-97.3% of the total sequences), Bacteroidetes (0.1-62.6%), Actinobacteria (0.2-14.7%), Fusobacteria (0.0-5.7%), and Proteobacteria (0.0-5.1%). The relative abundance of Firmicutes was significantly lower in the Maltese dog breed than that in the other two breeds, while that of Fusobacteria was significantly higher in the Maltese than in the Miniature Schnauzer breed. At the genus level, the relative abundance of Streptococcus, Fusobacterium, Turicibacter, Succinivibrio, and Anaerobiospirillum differed significantly among the three dog breeds. These genera had no correlation with age, diet, sex, body weight, vaccination history, or parasite protection history. Within a breed, some of these genera had a correlation with at least one blood chemistry value. This study indicates that the composition of the fecal microbiome in dogs is affected by breed.

Journal ArticleDOI
TL;DR: The use of nanotechnology is demonstrated to modify existing clinically-approved drugs and enhance their efficacy against pathogenic amoebae and this is a viable approach in the treatment of neglected diseases.
Abstract: Parasitic infections have remained a significant burden on human and animal health. In part, this is due to lack of clinically-approved, novel antimicrobials and a lack of interest by the pharmaceutical industry. An alternative approach is to modify existing clinically-approved drugs for efficient delivery formulations to ensure minimum inhibitory concentration is achieved at the target site. Nanotechnology offers the potential to enhance the therapeutic efficacy of drugs through modification of nanoparticles with ligands. Amphotericin B, nystatin, and fluconazole are clinically available drugs in the treatment of amoebal and fungal infections. These drugs were conjugated with gold nanoparticles. To characterize these gold-conjugated drug, atomic force microscopy, ultraviolet-visible spectrophotometry and Fourier transform infrared spectroscopy were performed. These drugs and their gold nanoconjugates were examined for antimicrobial activity against the protist pathogen, Acanthamoeba castellanii of the T4 genotype. Moreover, host cell cytotoxicity assays were accomplished. Cytotoxicity of these drugs and drug-conjugated gold nanoparticles was also determined by lactate dehydrogenase assay. Gold nanoparticles conjugation resulted in enhanced bioactivity of all three drugs with amphotericin B producing the most significant effects against Acanthamoeba castellanii (p < 0.05). In contrast, bare gold nanoparticles did not exhibit antimicrobial potency. Furthermore, amoebae treated with drugs-conjugated gold nanoparticles showed reduced cytotoxicity against HeLa cells. In this report, we demonstrated the use of nanotechnology to modify existing clinically-approved drugs and enhance their efficacy against pathogenic amoebae. Given the lack of development of novel drugs, this is a viable approach in the treatment of neglected diseases.

Journal ArticleDOI
TL;DR: The findings suggest that the dietary supplementation of β-1,3/1,6-glucan and/or L. plantarum LM1004 has a great potential for treatment of AD as well as obesity in humans through mechanisms that might involve modulation of host immune systems and gut microbiota.
Abstract: Atopic dermatitis (AD) is a chronic inflammatory skin disease of mainly infants and children. Currently, the development of safe and effective treatments for AD is urgently required. The present study was conducted to investigate the immunomodulatory effects of yeast-extracted β-1,3/1,6-glucan and/or Lactobacillus plantarum (L. plantarum) LM1004 against AD-like symptoms. To purpose, β-1,3/1,6-glucan and/or L. plantarum LM1004 were orally administered to AD-induced animal models of rat (histamine-induced vasodilation) and mouse (pruritus and contact dermatitis) exhibiting different symptoms of AD. We then investigated the treatment effects on AD-like symptoms, gene expression of immune-related factors, and gut microbiomes. Oral administration of β-1,3/1,6-glucan (0.01 g/kg initial body weight) and/or 2 × 1012 cells/g L. plantarum LM1004 (0.01 g/kg initial body weight) to ADinduced animal models showed significantly reduced vasodilation in the rat model, and pruritus, edema, and serum histamine in the mouse models (p < 0.05). Interestingly, β-1,3/1,6- glucan and/or L. plantarum LM1004 significantly decreased the mRNA levels of Th2 and Th17 cell transcription factors, while the transcription factors of Th1 and Treg cells, galactin-9, filaggrin increased, which are indicative of enhanced immunomodulation (p < 0.05). Moreover, in rats with no AD induction, the same treatments significantly increased the relative abundance of phylum Bacteroidetes and the genus Bacteroides. Furthermore, bacterial taxa associated with butyrate production such as, Lachnospiraceae and Ruminococcaceae at family, and Roseburia at genus level were increased in the treated groups. These findings suggest that the dietary supplementation of β-1,3/1,6-glucan and/or L. plantarum LM1004 has a great potential for treatment of AD as well as obesity in humans through mechanisms that might involve modulation of host immune systems and gut microbiota.

Journal ArticleDOI
TL;DR: An overview of the latest developments in BSSD technology and its application in protein engineering is presented.
Abstract: Bacillus subtilis spore surface display (BSSD) technology is considered to be one of the most promising approaches for expressing heterologous proteins with high activity and stability. Currently, this technology is used for various purposes, such as the production of enzymes, oral vaccines, drugs and multimeric proteins, and the control of environmental pollution. This paper presents an overview of the latest developments in BSSD technology and its application in protein engineering. Finally, the major limitations of this technology and future directions for its research are discussed.

Journal ArticleDOI
TL;DR: This study aimed to determine the modulations of the IFN promoter activity, induced by the MDA5/RIG-I signaling pathway, by over-expressing individual ZIKV genes and showed that two nonstructural proteins, NS2A and NS4A, significantly down-regulated the promoter activity of IFn-β by inhibiting multiple signaling molecules involved in the activation of IFN-β.
Abstract: Zika virus (ZIKV) is a mosquito-transmitted, emerging Flavivirus that causes Guillain-Barre syndrome and microcephaly in adults and fetuses, respectively. Since ZIKV was first isolated in 1947, severe outbreaks have occurred at various places worldwide, including Yap Island in 2007, French Polynesia in 2013, and Brazil in 2015. Although incidences of ZIKV infection and dissemination have drastically increased, the mechanisms underlying the pathogenesis of ZIKV have not been sufficiently studied. In addition, despite extensive research, the exact roles of individual ZIKV genes in the viral evasion of the host innate immune responses remain elusive. Besides, it is still possible that more than one ZIKV-encoded protein may negatively affect type I interferon (IFN) induction. Hence, in this study, we aimed to determine the modulations of the IFN promoter activity, induced by the MDA5/RIG-I signaling pathway, by over-expressing individual ZIKV genes. Our results show that two nonstructural proteins, NS2A and NS4A, significantly down-regulated the promoter activity of IFN-β by inhibiting multiple signaling molecules involved in the activation of IFN-β. Interestingly, while NS2A suppressed both full-length and constitutively active RIG-I, NS4A had inhibitory activity only on full-length RIG-I. In addition, while NS2A inhibited all forms of IRF3 (full-length, regulatory domain-deficient, and constitutively active), NS4A could not inhibit constitutively active IRF3-5D. Taken together, our results showed that NS2A and NS4A play major roles as antagonists of MDA5/RIG-I-mediated IFN-β induction and more importantly, these two viral proteins seem to inhibit induction of the type I IFN responses in differential mechanisms. We believe this study expands our understanding regarding the mechanisms via which ZIKV controls the innate immune responses in cells and may pave the way to development of ZIKV-specific therapeutics.

Journal ArticleDOI
TL;DR: Results suggest that noble L. brevis KCCM 12203P could be used as a probiotic or applied to functional food processing and pharmaceutical fields for immunocompromised people.
Abstract: The purpose of this study was to determine the probiotic properties of Lactobacillus brevis KCCM 12203P isolated from the Korean traditional food kimchi and to evaluate the antioxidative activity and immune-stimulating potential of its heat-killed cells to improve their bio-functional activities. Lactobacillus rhamnosus GG, which is a representative commercial probiotic, was used as a comparative sample. Regarding probiotic properties, L. brevis KCCM 12203P was resistant to 0.3% pepsin with a pH of 2.5 for 3 h and 0.3% oxgall solution for 24 h, having approximately a 99% survival rate. It also showed strong adhesion activity (6.84%) onto HT-29 cells and did not produce β-glucuronidase but produced high quantities of leucine arylamidase, valine arylamidase, β-galactosidase, and N-acetyl-β- glucosaminidase. For antioxidant activity, it appeared that viable cells had higher radical scavenging activity in the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assay, while in the 2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assay, heat-killed cells had higher antioxidant activity. Additionally, L. brevis KCCM 12203P showed higher lipid oxidation inhibition ability than L. rhamnosus GG; however, there was no significant difference (p < 0.05) between heat-killed cells and control cells. Furthermore, heat-killed L. brevis KCCM 12203P activated RAW 264.7 macrophage cells without cytotoxicity at a concentration lower than 108 CFU/ml and promoted higher gene expression levels of inducible nitric oxide synthase, interleukin-1β, and interleukin-6 than L. rhamnosus GG. These results suggest that novel L. brevis KCCM 12203P could be used as a probiotic or applied to functional food processing and pharmaceutical fields for immunocompromised people.

Journal ArticleDOI
TL;DR: Investigation of the immune-stimulatory function of heat-killed Lactobacillus plantarum KCTC 13314BP indicates that LBP can be a promising immune-enhancing functional food agent.
Abstract: Identification of novel probiotic strains is of great interest in the field of functional foods. Specific strains of heat-killed bacteria have been reported to exert immunomodulatory effects. Herein, we investigated the immune-stimulatory function of heat-killed Lactobacillus plantarum KCTC 13314BP (LBP). Treatment with LBP significantly increased the production of TNF-α and IL-6 by macrophages. More importantly, LBP was able to enhance the phagocytic activity of macrophages against bacterial particles. Activation of p38, JNK, ERK, NF-κB, and STAT3 was involved in the immunomodulatory function of LBP. LBP treatment significantly increased production of TNF-α by bone marrow-derived macrophages and splenocytes, further confirming the immunostimulatory effect of LBP in primary immune cells. Interestingly, the immunomodulatory effects of LBP were much stronger than those of Lactobacillus rhamnosus GG, a well-known probiotic strain. These results indicate that LBP can be a promising immune-enhancing functional food agent.

Journal ArticleDOI
TL;DR: Results proved the potential of lipid production from A. donax, which is particularly important for their consideration as substitutes for vegetable oils in many applications such as biodiesel or bioplastics.
Abstract: The use of low-cost substrates represents one key issue to make single cell oil production sustainable. Among low-input crops, Arundo donax L. is a perennial herbaceous rhizomatous grass containing both C5 and C6 carbohydrates. The scope of the present work was to investigate and optimize the production of lipids by the oleaginous yeast Cutaneotrichosporon curvatus from undetoxified lignocellulosic hydrolysates of steam-pretreated A. donax. The growth of C. curvatus was first optimized in synthetic media, similar in terms of sugar concentration to hydrolysates, by applying the response surface methodology (RSM) analysis. Then the bioconversion of undetoxified hydrolysates was investigated. A fed-batch process for the fermentation of A. donax hydrolysates was finally implemented in a 2-L bioreactor. Under optimized conditions, the total lipid content was 64% of the dry cell weight and the lipid yield was 63% of the theoretical. The fatty acid profile of C. curvatus triglycerides contained 27% palmitic acid, 33% oleic acid and 32% linoleic acid. These results proved the potential of lipid production from A. donax, which is particularly important for their consideration as substitutes for vegetable oils in many applications such as biodiesel or bioplastics.

Journal ArticleDOI
TL;DR: The endophytic bacteria promoted in plant growth and yield under drought had outstanding function in the enhancement of inulin content under well-watered condition.
Abstract: Drought is more concerned to be a huge problem for agriculture as it affects plant growth and yield. Endophytic bacteria act as plant growth promoting bacteria that have roles for improving plant growth under stress conditions. The properties of four strains of endophytic bacteria were determined under water deficit medium with 20% polyethylene glycol. Bacillus aquimaris strain 3.13 showed high 1-aminocyclopropane-1-carboxylate (ACC) deaminase production; Micrococcus luteus strain 4.43 produced indole acetic acid (IAA). Exopolysaccharide production was high in Bacillus methylotrophicus strain 5.18 while Bacillus sp. strain 5.2 did not show major properties for drought response. Inoculation of endophytic bacteria into plants, strain 3.13 and 4.43 increased height, shoot and root weight, root length, root diameter, root volume, root area and root surface of Jerusalem artichoke grown under water limitation, clearly shown in water supply at 1/3 of available water. These increases were caused by bacteria ACC deaminase and IAA production; moreover, strain 4.43 boosted leaf area and chlorophyll levels, leading to increased photosynthesis under drought at 60 days of planting. The harvest index was high in the treatment with strain 4.43 and 3.13 under 1/3 of available water, promoting tuber numbers and tuber weight. Inulin content was unchanged in the control between well-watered and drought conditions. In comparison, inulin levels were higher in the endophytic bacteria treatment under both conditions, although yields dipped under drought. Thus, the endophytic bacteria promoted in plant growth and yield under drought; they had outstanding function in the enhancement of inulin content under wellwatered condition.

Journal ArticleDOI
TL;DR: It was found that acetic acid in the undissociated state can diffuse freely through the hydrophobic layer of the membrane and dissociate, affecting the production, yield, and activity of bacteriocin.
Abstract: Acetic acid is indirectly involved in cell center metabolism, and acetic acid metabolism is the core of central metabolism, affecting and regulating the production of bacteriocin. Bacteriocin is a natural food preservative that has been used in the meat and dairy industries and winemaking. In this paper, the effects of acetic acid on bacteriocin produced by Gram-positive bacteria were reviewed. It was found that acetic acid in the undissociated state can diffuse freely through the hydrophobic layer of the membrane and dissociate, affecting the production, yield, and activity of bacteriocin. In particular, the effect of acetic acid on cell membranes is summarized. The link between acetic acid metabolism, quorum sensing, and bacteriocin production mechanisms is also highlighted.

Journal ArticleDOI
TL;DR: The molecular and physiological characteristics of the three types of programmed cell death and their development potential as an antibacterial agent are addressed.
Abstract: In the bacterial community, unicellular organisms act together as a multicellular being. Bacteria interact within the community and programmed cell death (PCD) in prokaryotes is a sort of altruistic action that enables the whole population to thrive. Genetically, encoded cell death pathways are triggered by DNA damage or nutrient starvation. Given the environmental and bacterial diversity, different PCD mechanisms are operated. Still, their biochemical and physiological aspects remain unrevealed. There are three main pathways; thymineless death, apoptosis-like death, and toxin-antitoxin systems. The discovery of PCD in bacteria has revealed the possibility of developing new antibiotics. In this review, the molecular and physiological characteristics of the three types of PCD and their development potential as antibacterial agents are addressed.

Journal ArticleDOI
TL;DR: This review summarizes the biochemical data for the two major metalloregulator families, SmtB/ArsR and MerR, to describe their metal-binding sites, specific chelating chemistry, and conformational changes.
Abstract: To adapt to environmental changes and to maintain cellular homeostasis, microorganisms adjust the intracellular concentrations of biochemical compounds, including metal ions; these are essential for the catalytic function of many enzymes in cells, but excessive amounts of essential metals and heavy metals cause cellular damage. Metal-responsive transcriptional regulators play pivotal roles in metal uptake, pumping out, sequestration, and oxidation or reduction to a less toxic status via regulating the expression of the detoxification-related genes. The sensory and regulatory functions of the metalloregulators have made them as attractive biological parts for synthetic biology, and the exceptional sensitivity and selectivity of metalloregulators toward metal ions have been used in heavy metal biosensors to cope with prevalent heavy metal contamination. Due to their importance, substantial efforts have been made to characterize heavy metal-responsive transcriptional regulators and to develop heavy metal-sensing biosensors. In this review, we summarize the biochemical data for the two major metalloregulator families, SmtB/ArsR and MerR, to describe their metal-binding sites, specific chelating chemistry, and conformational changes. Based on our understanding of the regulatory mechanisms, previously developed metal biosensors are examined to point out their limitations, such as high background noise and a lack of well-characterized biological parts. We discuss several strategies to improve the functionality of the metal biosensors, such as reducing the background noise and amplifying the output signal. From the perspective of making heavy metal biosensors, we suggest that the characterization of novel metalloregulators and the fabrication of exquisitely designed genetic circuits will be required.