An, Jeong-Lib;Kim, Sang-Ha;Yu, Young-Bin;Kim, Sunghyun;Lee, Moo-Sik;Kim, Young-Kwon
Biomedical Science Letters
/
v.27
no.4
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pp.264-269
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2021
The purpose of this study was to evaluate the disinfecting efficacy of peracetic acid (PAA), sodium hypochlorite (NaOCl) and phenol, which are representative disinfectants in medical environments using four types of multi-drug resistance (MDR) clinical isolates with healthcare-associated infections (HAI). 26 antibiotic susceptibility tests were conducted for the four types of MDR clinical isolates in the same way as for clinical specimens. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the disinfectants were determined by using in vitro liquid medium dilution method and inoculation of the plate medium. Both the MIC and MBC of phenol against MRSA and VRE were 3.1%, while those against KPC and MRPA were 6.2%. The MIC and MBC of peracetic acid (PAA) against MRSA, VRE, KPC, and MRPA were 0.18%. The MIC and MBC of sodium hypochlorite (NaOCl) against MRSA were 0.39% and 0.78%, respectively. Both values of MIC and MBC were 0.78% for VRE. In addition, KPC and MRPA showed 0.39% for MIC and 0.78% for MBC. For all MDR strains used in this study, sodium hypochlorite and peracetic acid showed significant sterilizing efficiency, while no clear correlation was identified between antibiotic resistance clinical isolated and ability of disinfection.
Continued fenvalerate use has caused serious environmental pollution and requires large-scale remediation. Dibutyl phthalate (DBP) was discovered in fenvalerate metabolites degraded by Citrobacter freundii CD-9. Coculturing is an effective method for bioremediation, but few studies have analyzed the degradation pathways and potential mechanisms of cocultures. Here, a DBP-degrading strain (BDBP 071) was isolated from soil contaminated with pyrethroid pesticides (PPs) and identified as Stenotrophomonas acidaminiphila. The optimum conditions for DBP degradation were determined by response surface methodology (RSM) analysis to be 30.9 mg/l DBP concentration, pH 7.5, at a culture temperature of 37.2℃. Under the optimized conditions, approximately 88% of DBP was degraded within 48 h and five metabolites were detected. Coculturing C. freundii CD-9 and S. acidaminiphila BDBP 071 promoted fenvalerate degradation. When CD-9 was cultured for 16 h before adding BDBP 071, the strain inoculation ratio was 5:5 (v/v), fenvalerate concentration was 75.0 mg/l, fenvalerate was degraded to 84.37 ± 1.25%, and DBP level was reduced by 5.21 mg/l. In addition, 12 fenvalerate metabolites were identified and a pathway for fenvalerate degradation by the cocultured strains was proposed. These results provide theoretical data for further exploration of the mechanisms used by this coculture system to degrade fenvalerate and DBP, and also offer a promising method for effective bioremediation of PPs and their related metabolites in polluted environments.
Mi-Hyun Lee;Sung-Jun Hong;Dong Suk Park;Hyeonheui Ham;Hyun Gi Kong
The Plant Pathology Journal
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v.39
no.4
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pp.409-416
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2023
Bacterial leaf blight of carrots caused by Xanthomonas hortorum pv. carotae (Xhc) is an important worldwide seed-borne disease. In 2012 and 2013, symptoms similar to bacterial leaf blight were found in carrot farms in Jeju Island, Korea. The phenotypic characteristics of the Korean isolation strains were similar to the type strain of Xhc. Pathogenicity showed symptoms on the 14th day after inoculation on carrot plants. Identification by genetic method was multi-position sequencing of the isolated strain JJ2001 was performed using four genes (danK, gyrB, fyuA, and rpoD). The isolated strain was confirmed to be most similar to Xhc M081. Furthermore, in order to analyze the genetic characteristics of the isolated strain, whole genome analysis was performed through the next-generation sequencing method. The draft genome size of JJ2001 is 5,443,372 bp, which contains 63.57% of G + C and has 4,547 open reading frames. Specifically, the classification of pathovar can be confirmed to be similar to that of the host lineage. Plant pathogenic factors and determinants of the majority of the secretion system are conserved in strain JJ2001. This genetic information enables detailed comparative analysis in the pathovar stage of pathogenic bacteria. Furthermore, these findings provide basic data for the distribution and diagnosis of Xanthomonas hortorum pv. carotae, a major plant pathogen that infects carrots in Korea.
The productions of beta-exotoxin from sixteen Bacillus thuringiensis strains were examined by Micrococus flava primarily, and then measured by spectrophotometer during culturing in Conner and Hansen mineral salts medium at 28$^{\circ}C$. Also the toxic effects of the toxin to mice were checked. The growth of Bacillus thuringiensis K2 and BTK2-T1, -T13, -T33 and -T40 got into stationary phase at 6 hour culture and then maintained it up to 48 hours without severe fluctuation. The production of beta-exotoxin from the strains, BTK2, BTK2-T1, -T13, -T17 and -T33 appeared at 6 hour culture and the amounts of the toxin were about 40 $\mu\textrm{g}$/$m\ell$ at 6 hour culture, approximately 70 $\mu\textrm{g}$/$m\ell$ at 12 hours, approximately 85$\mu\textrm{g}$/$m\ell$ from 24 hours to 48 hours. At 48 hour-culture, BTK2 produced 80 $\mu\textrm{g}$/$m\ell$ of beta-exotoxin (5.5$\times$10$^{8}$ cells/$m\ell$, BTK2-T13 produced 84 $\mu\textrm{g}$/$m\ell$ (4.3$\times$10$^{8}$ cells/$m\ell$), BTK2-T17 produced 87$\mu\textrm{g}$/$m\ell$ (1.4$\times$10$^{8}$ cells/$m\ell$), and BTK2-T33 produced 84 $\mu\textrm{g}$/$m\ell$ (4.9$\times$10$^{8}$ cells/$m\ell$). All other serotypes also produced beta-exotoxin. At 48 hour culture, BTK-37 produced 88$\mu\textrm{g}$/$m\ell$ (6.1$\times$10$^{8}$ cells/$m\ell$), BTK-35 produced 81 $\mu\textrm{g}$/$m\ell$), and the rest of them produced less than 70 $\mu\textrm{g}$/$m\ell$. To check the toxicity of beta-exotoxin and B. thuringiensis, the cultured media with microorganisms were inoculated to mice by per os, intraperiloneal, subcutaneous and intracerebral injection, and nasal cavity inoculation for 30 days. However, the toxin did not kill all of the treated mice.
Kim, Hyeong-Hwan;Cho, Sung-Rae;Choo, Ho-Yul;Lee, Sang-Myeong;Jeon, Heung-Yong;Lee, Dong-Woon
Korean journal of applied entomology
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v.47
no.4
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pp.447-456
/
2008
Five effective strains against tobacco cutworm, Spodoptera litura (Lepidoptera: Noctuidae), Steinernema carpocapsae (GSN1), Steinernema sp. (GSNUS-10), Steinernema sp. (GSNUS-14), Heterorhabditis bacteriophora Hamyang (HbH), and Heterorhabditis sp. (GSNUH-1) were selected among 14 isolates of Korean entomopathogenic nematode in laboratory tests. $LC_{50}$ values of above five strains against tobacco cutworm were various by different nematode strains and developmental stages of tobacco cutworm. $LC_{50}$ value of S. carpocapsae (GSN1) was the lowest by $4.0{\sim}8.3$ infective juveniles (Ijs) and 2nd instars of tobacco cutworm was most susceptible. Pathogenicity of five effective strains against tobacco cutworm depends on nematode strain, concentration, and application times. The most effective strain was determined as S. carpocapsae (GSN1). Two or three times of applications were effective regardless of nematode strain, or concentration. Efficacy of S. carpocapsae (GSN1), Steinernema (GSNUS-10), Steinernema (GSNUS-14), and Heterorhabditis (GSNUH-1) was variable depending on nematode strain, concentration, application times, and host variety. S. carpocapsae (GSN1) was the most effective and inoculation of 100,000 infective juveniles per m2 (720,000 Ijs/7.2 $m^2=1{\times}10^9$ Ijs/ha) resulted in higher efficacy. Three times of application of nematodes led to higher control efficacy than one or two applications. Efficacy of nematodes was higher on Chinese cabbage than cabbage or kale.
Avian pathogenic Escherichia coli (APEC) causes a number of extraintestinal diseases in poultry. A virulence factor, P-fimbriae is firmly associated with the diseases. In this study, to develop an effective vaccine for the prevention of APEC, recombinant attenuatted Salmonella Typhimurium vaccines expressing PapA and PapG of P-fimbriae were evaluated whether these induced protective immune responses in murine models. Female BALB/c mice were primed and boosted orally at 7 and 10 weeks of age. In all immunized mice, the antigen-specific serum IgG levels were remained higher than those in the control mice from the fourth week post inoculation till the end of this study. In addition, antigen-specific serum IgG levels in the prime-booster immunized mice were enhanced as compared to the single immunized mice among each immunized group. The antigen-specific mucosal IgA levels in the mice immunized with each strain also induced higher than those in control mice. In addition, serum IgG and fecal IgA levels in mice administered with the combination of both strains were highly induced compared to those in mice immunized with each strain alone. These results indicated that PapA and PapG worked together for inducing high immune responses. To partly discern the nature of immunity induced by the strains, we quantified serum IgG subtypes IgG1 and IgG2a specific to antigens. The PapA and PapG strains biased the immunity to the Th1-type, as determined by the IgG2a/IgG1 ratio. On the other hand, the immunization with the both strains in combination produced mixed Th1- and Th2-type immune responses. These indicated that immunization with the combination of PapA and PapG could elicit both humoral and cell-mediated immunities.
Kim, Min-Jeong;Shim, Chang-Ki;Kim, Yong-Ki;Hong, Sung-Jun;Park, Jong-Ho;Han, Eun-Jung;Jee, Hyeong-Jin;Yun, Jong-Chul;Kim, Suk-Chul
Korean Journal of Organic Agriculture
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v.22
no.4
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pp.743-760
/
2014
This study aimed to isolate and identify freshwater algae from the organic agricultural ecosystems and investigate its biological characteristics to study the possibility of utilizing a biomass freshwater algae in organic farming. In the survey area, average water temperature was $12.4{\sim}28.2^{\circ}C$ and the pH ranges were from 6.1 to 8.5. The solid culture method is more suitable than liquid culture method for isolation of freshwater algae with lower contamination level and higher isolation frequency. A total of 115 strains were isolated from six freshwater algae habitats in nine regions in Korea. BGMM (BG11 Modified Medium) amended with NaNO3 and $KNO_3$ as a nitrogen, and $Na_2CO_3$ as carbon source was designed to isolate and culture freshwater algae. Absorbance of freshwater algae culture has increased dramatically to four days and decreased after eight days after inoculation. CHK008 of the seven isolates showed the highest absorbance in seven days after culturing in BGMM. The optimal pH of BGMM for culturing freshwater algae was pH 6-7. As light intensity increased, growth of freshwater algae increased. Among the five kinds of carbon sources, glucose and galactose promoted good growth of freshwater algae in BGMM. The colony color of purified 16 green algae isolates showed a separation of green, dark and light green, and of them, eleven algae strains showed a strong fluorescent light under fluorescence microscopy. Cell size of the green algae showed a wide range of variation depending on the species. General morphology of the green algae strains was spherical. Chlamydomonas sp. was elliptical, and Chlorella sorokiniana was ellipsoidal and cylindrical. All strains of the green algae except for Chlamydomonas sp. did not have flagella. One isolate of Chlamydomonas sp. and five isolates of C. sorokiniana secreted mucus. Sixteen isolates of 16 green algae were identified as two family and six species, Chlorella vulgalis, C. sorokiniana, C. pyrenoidosa, C. kessleri, C. emersonii, and Chlamydomonas sp. based on their morphological characteristics.
Infectious bursal disease (IBD) is an acute, highly contagious, and immunosuppressive disease in young chickens, and causes considerable economic losses to the poultry industry. More than 30 years ago, an antigenic variant IBDV (avIBDV) was reported in chicken farms in the United States. Recently, a novel avIBDV exhibited clear differences in molecular characteristics compared with previous variant strains. This study investigated the molecular characteristics of recently isolated avIBDV strains in Korea. Strains of avIBDV were confirmed by reverse transcription PCR (RT-PCR) and were propagated in 10-day-old specific-pathogen-free (SPF) embryonated chicken eggs through chorioallantoic membrane (CAM) inoculation. Multiple sequence alignment and phylogenetic analyses of hypervariable regions VP2 gene revealed that the strains originated from two different avIBDV lineages (G2a and G2d). In our results, we confirmed the co-existence and prevalence of avIBDV genogroup G2a and G2d in chicken farms. It is necessary to study the protective efficacy of current vaccines against avIBDVs.
Kim, Yiseul;Kim, Sang Yoon;An, Ju Hee;Sang, Mee Kyung;Weon, Hang-Yeon;Song, Jaekyeong
Microbiology and Biotechnology Letters
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v.46
no.3
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pp.253-260
/
2018
Beneficial microorganisms are widely used in the forestry, livestock, and, in particular, agricultural sectors to control soilborne diseases and promote plant growth. However, the industrial utilization of these microorganisms is very limited, mainly due to uncertainty concerning their ability to colonize and persist in soil. In this study, the survival of beneficial microorganisms in field soil microcosms was investigated for 13 days using quantitative PCR with B. subtilis group-specific primers. Bacterial community dynamics of the treated soils were analyzed using 16S ribosomal RNA (rRNA) gene amplicon sequencing on the Illumina MiSeq platform. The average 16S rRNA gene copy number per g dry soil of Bacillus spp. was $4.37{\times}10^6$ after treatment, which was 1,000 times higher than that of the control. The gene copy number was generally maintained for a week and was reduced thereafter, but remained 100 times higher than that of the control. Bacterial community analysis indicated that Acidobacteria ($26.3{\pm}0.9%$), Proteobacteria ($24.2{\pm}0.5%$), Chloroflexi ($11.1{\pm}0.4%$), and Actinobacteria ($9.7{\pm}2.5%$) were abundant phyla in both treated and non-treated soils. In the treated soils, the relative abundance of Actinobacteria was lower, whereas those of Bacteroidetes and Firmicutes were higher compared to the control. Differences in total relative abundances of operational taxonomic units belonging to several genera were observed between the treated and non-treated soils, suggesting that inoculation of soil with the Bacillus strains influenced the relative abundances of certain groups of bacteria and, therefore, the dynamics of resident bacterial communities. These changes in resident soil bacterial communities in response to inoculation of soil with beneficial Bacillus spp. provide important information for the use of beneficial microorganisms in soil for sustainable agriculture.
Two attenuated Cucumber mosaic virus (CMV) isolates, Paf-CMV and Rs2-CMV that had been selected from CMV isolates associated with satellite RNA (satRNA) were tested for cross-protection effect in pepper plants. The viruses selected as attenuated strains appeared to be identical serologically and physically to the challenge virus (Mf-CMV), but they were lower in the dilution end-point of infectivity of crude sap than Mf-CMV When symptoms were observed in several indicator plants after inoculation, Paf-CMV and Rs2-CMV were symptomless or showed mild mosaic symptoms while another satRNA isolate Ap-CMV developed severe mosaic symptoms on the leaves as Mf-CMV. The nucleotide sequences of the satRNAs were determined by sequencing full-length cDNA clones. Paf-, Rs2- and Ap-satRNAs were 386, 335, and 347 nucleotides long, respectively, The sequences were then compared with the other known Y-satRNA, revealing that nucleotide sequences of the satRNAs consisted of 5'- and 3'-terminal conserved regions. However variations occurred on the middle regions of the sequences, especially those related to symptom interference, showing significant differences between Paf-satRNA and other isolates. Infectious transcripts of Paf-satRNA and Rs2-satRNA induced mild mosaic symptoms in pepper plants when supported by genomic RNAs of Mf-CMV. Under greenhouse conditions, Paf-CMV and Rs2-CMV were tested for cross-protection effect in pepper and tobacco (Nicotiana tabacum cv, Xanthi nc) plants against Mf-CMV. No symptoms were developed on the plants vaccinated with Paf-CMV until 3 weeks after inoculation with the virulent strain; however another attenuated isolate, Rs2-CMV, showed less effectiveness in cross-protection. Depending on the concentration of the challenged virus, symptoms sometimes appeared later in the upper leaves. However, in plants challenged with low concentrations (below 0.2 mg/ml) of the challenge inoculum, symptoms caused by the virulent strain did not develop on the plants vaccinated with Paf-CMV. In the field experiments, the number of pepper plants with severe mosaic symptoms in the control plots was progressively increased after transplanting and reached approximately 50% after 50 days. On the other hand, the incidence of mosaic disease appeared very low on the plants that had received the protective inoculation with Paf-CMV.
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