A bacterial strain PN33 producing large amounts of extracellular pectin lyase (PNL, EC 4.2.2.10) was isolated from soil. The isolated bacterium was identified as a strain of Bacillus sp. Production of PNL by the strain was induced only by pectins, with a higher degree of esterification, which had been added to the culture medium as a sole carbon source. The optimal medium for PNL production was determined to consist of 10 g pectin, 2 g yeast extract, 4 g $K_2HPO_4{\cdot}3H_2O$, 0.6 g $MgSO_4$, and 0.11 g $CaCl_2$ per liter (pH 7.0). The PNL activity in the culture supernatant reached the highest level of 132 mU/ml after 32 h cultivation at $37^{\circ}C$ in the optimal medium. The PNL produced was purified to homogeneity by ammonium sulfate fractionation (50~80%), and cation exchange and size exclusion chromatographies. The molecular mass of the enzyme was estimated to be approximately 52 kDa by SDS-PAGE. Almost the same mass was determined by nondenaturing PAGE, indicating that the functional enzyme had a monomeric structure. As expected, the PNL exhibited higher activities on the highly esterified pectins whereas it gave no detectable activity on polygalacturonic acid. The enzyme showed the highest activity at the acidic pH of 6.0, exceptional for a bacterial PNL. Maximum activity was measured at $40^{\circ}C$, although the stability f the purified enzyme was poor at this temperature. alcium (1 mM) was found to activate the PNL activity by $50\%$, and also remarkably increased the thermal stability f the enzyme. Phenylmethylsulfonylfluoride (PMSF) and iethylpyrocarbonate (DEPC) inhibited the PNL activity lmost completely at the concentration of 5 mM. This result ndicates that some serine and histidine residues of the nzyme may play an essential role for catalytic function of he enzyme.
Two bacterial isolates obtained from rotifer and diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization. In an attempt to develop rapid PCR method for the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. Analysis of the ISR sequences showed that V. ichthyoenteri contains one type of polymorphic ISRs. The size of ISRs was 348 bp length and did not contain tRNA genes. Mutiple alignment of representative sequences from different V. species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of V. ichthyoenteri. The specificity of the primer was examined using genomic DNA prepared from 19 different V. species, isolated 18group Vibrio species and most similar sequence of other known Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.
Han, Gui Hwan;Bong, Ki Moon;Kim, Jong Min;Kim, Pyoung Il;Kim, Si Wouk
KSBB Journal
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v.29
no.3
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pp.131-138
/
2014
In this study, we isolated two novel chitinase producing bacterial strains from yellow loess samples collected from Jullanamdo province. The chitinase producing bacteria were isolated based on the zone size of clearance in the chitin agar plates. Both of them were gram positive, rod ($2{\sim}3{\times}0.3{\sim}0.4{\mu}m$), spore-forming, and motility positive. They were facultative anaerobic, catalase positive and hydrolyzed starch, gelatin, and casein. From the 16s rRNA gene sequence analysis, the isolates were labeled as Bacillus licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02. The isolates showed higher extracellular chitinase activities than B. licheniformis ATCC 14580 as a control. The optimum temperature and pH for chitinase production were $40^{\circ}C$ and pH 7.0, respectively. Response Surface Methodology (RSM) was used to optimize the culture medium for efficient production of the chitinase. Under this optimal condition, 1.5 times higher chitinase activity of B. licheniformis KYLS-CU02 was obtained. Extracellular chitinases of the two isolates were purified through ammonium sulfate precipitation and anion-exchange DEAE-cellulose column chromatography. The specific activities of purified chitinase from B. licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02 were 7.65 and 5.21 U/mg protein, respectively. The molecular weights of the two purified chitinases were 59 kDa. Further, the purified chitinase of B. licheniformis KYLS-CU01 showed high antifungal activity against Fusarium sp.. In conclusion, these two bacterial isolates can be used as a biopesticide to control pathogenic fungi.
Biological control agents (BCAs) from different microbial taxa are increasingly used to control bacterial wilt caused by Ralstonia solanacearum. However, a quantitative research synthesis has not been conducted on the role of BCAs in disease suppression. Therefore, the present study aimed to meta-analyze the impacts of BCAs on both Ralstonia wilt disease suppression and plant (host) growth promotion. The analysis showed that the extent of disease suppression by BCAs varied widely among studies, with effect size (log response ratio) ranging from -2.84 to 2.13. The disease incidence and severity were significantly decreased on average by 53.7% and 49.3%, respectively. BCAs inoculation also significantly increased fresh and dry weight by 34.4% and 36.1%, respectively on average. Also, BCAs inoculation significantly increased plant yield by 66%. Mean effect sizes for genus Pseudomonas sp. as BCAs were higher than for genus Bacillus spp. Among antagonists tested, P. fluorescens, P. putida, B. cereus, B. subtilis and B. amyloliquefaciens were found to be more effective in general for disease reduction. Across studies, highest disease control was found for P. fluorescens, annual plants, co-inoculation with more than one BCA, soil drench and greenhouse condition were found to be essential in understanding plant responses to R. solanacearum. Our results suggest that more efforts should be devoted to harnessing the potential beneficial effects of these antagonists, not just for plant growth promoting traits but also in mode of applications, BCAs formulations and their field studies should be considered in the future for R. solanacearum wilt disease suppression.
DFA IV is di-D-fructose-2,6':6,2'-dianhydride, consisting of two fructose residues. It can be enzymatically synthesized from levan by levan fructotransferase, and can be used for mineral absorption. Understanding of the structure and composition of levan is important to obtain high-level production of DFA IV. A bacterial strain, Pseudomonas aurantiaca 5-4380, was identified to produce low-branched levan, and the levansucrase gene (lsch) from this bacterium was found to be composed of 1,275 Up coding for a protein of 424 amino acids, with an estimated molecular weight of 47 kDa. The bacterial levansucrase gene was expressed in Escherichia coli DH5${\alpha}$ by its own promoter and lac promoter. The recombinant levansucrase was produced in soluble form with 170U of levansucrase activity from 1-ml E. coii culture broth. The expressed enzyme from the clone showed similar biochemical properties, such as size of active levansucrase, degree of branching, and optimum temperature, with P.aurantiaca 5-4380 levansucrase.
Kim, Grace Yoon-Hee;Hong, Chang-Wan;Park, Jung-Hyun
IMMUNE NETWORK
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v.11
no.1
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pp.1-10
/
2011
Interleukin-7 (IL-7) is an essential cytokine for T cells. However, IL-7 is not produced by T cells themselves such that T cells are dependent on extrinsic IL-7. In fact, in the absence of IL-7, T cell development in the thymus as well as survival of naive T cells in the periphery is severely impaired. Furthermore, modulating IL-7 availability in vivo either by genetic means or other experimental approaches determines the size, composition and function of the T cell pool. Consequently, understanding IL-7 expression is critical for understanding T cell immunity. Until most recently, however, the spatiotemporal expression of in vivo IL-7 has remained obscured. Shortage of such information was partly due to scarce expression of IL-7 itself but mainly due to the lack of adequate reagents to monitor IL-7 expression in vivo. This situation dramatically changed with a recent rush of four independent studies that describe the generation and characterization of IL-7 reporter mice, all utilizing bacterial artificial chromosome transgene technology. The emerging consensus of these studies confirmed thymic stromal cells as the major producers of IL-7 but also identified IL-7 reporter activities in various peripheral tissues including skin, intestine and lymph nodes. Strikingly, developmental and environmental cues actively modulated IL-7 reporter activities in vivo suggesting that IL-7 regulation might be a new mechanism of shaping T cell development and homeostasis. Collectively, the availability of these new tools opens up new venues to assess unanswered questions in IL-7 biology in T cells and beyond.
In order to understand the temporal distribution of pico- and nanoplankton and factors controlling its distribution at a station in Okkye Bay of Masan Bay located in the southern part of Korea, this study was conducted on two weeks interval from April 2005 to April 2006, and several abiotic and biotic factors were measured. During the study, picoplankton consisted of picoflagellates, cyanobacteria and heterotrophic bacteria, and nanoplankton consisted of nanoflagellates excluding dinoflagellates. The concentration of chlorophyll-a (chl-a) was a mean of $4.33\;{\mu}g/L$, and the nanoplanktonic ($<20\;{\mu}m$) chl-a size fraction was a mean of 39.5 % and significantly correlated with water temperature. The abundances of cyanobacteria and photosynthetic flagellates (PF) were means of $24.4{\times}10^{3}\;cells/mL\;and\;2.87{\times}10^{3}\;cells/mL$, respectively. The contribution of picoflagellates to the PF abundance varied among the sampling occasions and was a mean of 29 %, but to the PF carbon biomass was 2.6 % only. The PF abundance had significant relationships with water temperature, and silicate and TIN concentrations, suggesting that the PF abundance seemed to be primarily bottom-up regulated. The abundance of heterotrophic bacteria was a mean of $3.18{\times}10^{6}\;cells/mL$ and unlike other ecosystems it did not have relationships with chl-a and heterotrophic flagellates (HF), suggesting that bacterial abundance did not seem to be bottom-up or top-down regulated. HF mostly consisted of cells less than $5{\mu}m$ and its abundance was a mean of $2.71{\times}10^{3}\;cells/mL$. Of the HF abundance, picoflagellates occupied about 31 %, and occupied about 9 % of the HF carbon biomass. HF grazing activity on heterotrophic bacteria was relatively low and removed about 10 % of bacterial abundance, suggesting that HF might not be major consumers of bacteria and there seems to be other consumers in Okkye Bay. These results suggest that Okkye Bay may have a unique microbial ecosystem.
This case describes outbreaks of acute aspergillosis in a red-crowned crane. A six-month-old, male, crane had showed clinical signs (i.e. anorexia, performance loss, ruffled feathers and drooped wings and open mouth breathing, etc.) before death. In necropsy examination, spherical to oval nodules disseminated from the respiratory tract to other organs. Those nodules were formed predominantly in air sacs, lung, peritoneum, serosa of esophagus and trachea. The nodules varied in size from 1 mm to over 1cm and the color was white to yellow. Microscopically, most of lung architecture were replaced by multiple foci which were characterized by well demarcated eosinophilic and karyorrhetic debris and surrounded by numerous Inflammatory cell. Most within necrotic center of the nodules, large numbers of fungal hyphae were present. Microbiology result indicated fungal growths on sabroud dextrose agar and bacterial growths on blood agar. Bacteria identified as E. rhusiopathiae using MALDI-TOF (microflex, BRUKER, USA) and fungi identified as A. fumigatus, A. terreus by sequencing the ITS1 and ITS4 regions. To confirm the route of infection, we checked the existence of the same pathogens in cohabitant (i.e. mother crane). The young age and weakened immunity (i.e. bacterial infection, etc.) causes fatal aspergillosis in birds.
Objective : The treatment of choice for spinal epidural abscess (SEA) generally is urgent surgery in combination with intravenous antibiotic treatment. However, the optimal duration of antibiotic treatment has not been established to date, although 4-8 weeks is generally advised. Moreover, some researchers have reported that bacteremia is a risk factor for failure of antibiotic treatment in SEA. In this study, we investigated the clinical characteristics of SEA accompanied by bacteremia and also determined whether the conventional 4-8 weeks of antibiotic treatment is sufficient. Methods : We retrospectively reviewed the medical records and radiological data of 23 patients with bacterial SEA who underwent open surgery from March 2010 to April 2020. All patients had bacteremia preoperatively and underwent weeks of perioperative antibiotic treatments based on their identified organisms until all symptoms of infection disappeared. All patients underwent microbiological studies of peripheral blood, specimens from SEA and concomitant infections. The mean follow-up duration was 35.2 months, excluding three patients who died. Results : The male : female ratio was 15 : 8, and the mean age was 68.9 years. The SEA most commonly involved the lumbar spinal segment (73.9%), and the mean size was 2.9 vertebral body lengths. Mean time periods of 8.4 days and 16.6 days were required from admission to diagnosis and from admission to surgery, respectively. Concomitant infections more frequently resulted in delayed diagnosis (p=0.032), masking the symptoms of SEA. Methicillin-sensitive Staphylococcus aureus was the most commonly identified pathogen in both blood and surgical specimens. Seventeen patients (73.9%) showed no deficits at the final follow-up. The overall antibiotic treatment duration was a mean of 66.6 days, excluding three patients who died. This duration was longer than the conventionally advised 4-8 weeks (p=0.010), and psoas or paraspinal abscess required prolonged duration of antibiotic treatment (p=0.038). Conclusion : SEA accompanied by bacteremia required a longer duration (>8 weeks) of antibiotic treatment. In addition, the diagnosis was more frequently delayed in patients with concomitant infections. The duration of antibiotic treatment should be extended for SEA with bacteremia, and a high index of suspicion is mandatory for early diagnosis, especially in patients with concomitant infections.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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v.19
no.1
/
pp.1-10
/
2006
This experimental study was performed to investigate the effect of herbal eye drops, Tangpo-san and Coptidis rhizoma on Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa keratitis. The following results were obtained by using Minimum inhibition Concentration(MIC) and inhibition Zone. 1. MIC on Staphylococcus aureus in Tangpo-san was 100%, in Coptidis rhizoma was 100% and in Cravit was 0.1% 2. MIC on Staphylococcus epidermidis in Tangpo-san was 100%, in Coptidis rhizoma was 10% and in Cravit was 0.1%. 3. MIC on Pseudomonas aeruginosa in Tangpo-san, Coptidis rhizoma was not showing and in Cravit was 0.1%. 4. The size of inhibition zone on Staphylococcus aureus for Tangpo-san was 13.3mm in $50{\mu}{\ell}$, for Coptidis rhizoma was 26mm in $50{\mu}{\ell}$ and for Cravit was 31mm in $50{\mu}{\ell}$, showing the highest antibacterial effect. 5. The size of inhibition zone on Staphylococcus epidermidis for Tangpo-san was 16mm in $50{\mu}{\ell}$, for Coptidis rhizoma was 25mm in $40{\mu}{\ell}$ and for Cravit was 34mm in $50{\mu}{\ell}$, showing the highest antibacterial effect. 6. The size of inhibition zone on Pseudomonas aeruginosa for Tangpo-san, Coptidis rhizoma was not and for Cravit was 24.7mm in $50{\mu}l$, showing the antibacterial effect. In addition, the results shows that the herbal eye drops, Tangpo-san and Coptidis rhizoma can be used to cure Staphylococcus aureus, Staphylococcus epidermidis keratitis and if further study is performed, the use of herbal eye drops will be valuable and beneficial in the clinical medicines.
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