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RNA-seq Gene Profiling Reveals Transcriptional Changes in the Late Phase during Compatible Interaction between a Korean Soybean Cultivar (Glycine max cv. Kwangan) and Pseudomonas syringae pv. syringae B728a

  • Myoungsub, Kim;Dohui, Lee;Hyun Suk, Cho;Young-Soo, Chung;Hee Jin, Park;Ho Won, Jung
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.603-615
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    • 2022
  • Soybean (Glycine max (L) Merr.) provides plant-derived proteins, soy vegetable oils, and various beneficial metabolites to humans and livestock. The importance of soybean is highly underlined, especially when carbon-negative sustainable agriculture is noticeable. However, many diseases by pests and pathogens threaten sustainable soybean production. Therefore, understanding molecular interaction between diverse cultivated varieties and pathogens is essential to developing disease-resistant soybean plants. Here, we established a pathosystem of the Korean domestic cultivar Kwangan against Pseudomonas syringae pv. syringae B728a. This bacterial strain caused apparent disease symptoms and grew well in trifoliate leaves of soybean plants. To examine the disease susceptibility of the cultivar, we analyzed transcriptional changes in soybean leaves on day 5 after P. syringae pv. syringae B728a infection. About 8,900 and 7,780 differentially expressed genes (DEGs) were identified in this study, and significant proportions of DEGs were engaged in various primary and secondary metabolisms. On the other hand, soybean orthologs to well-known plant immune-related genes, especially in plant hormone signal transduction, mitogen-activated protein kinase signaling, and plant-pathogen interaction, were mainly reduced in transcript levels at 5 days post inoculation. These findings present the feature of the compatible interaction between cultivar Kwangan and P. syringae pv. syringae B728a, as a hemibiotroph, at the late infection phase. Collectively, we propose that P. syringae pv. syringae B728a successfully inhibits plant immune response in susceptible plants and deregulates host metabolic processes for their colonization and proliferation, whereas host plants employ diverse metabolites to protect themselves against infection with the hemibiotrophic pathogen at the late infection phase.

Chemical Composition and Quorum Sensing Inhibitory Effect of Nepeta curviflora Methanolic Extract against ESBL Pseudomonas aeruginosa

  • Haitham Qaralleh
    • Journal of Pharmacopuncture
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    • v.26 no.4
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    • pp.307-318
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    • 2023
  • Objectives: Bacterial biofilm is regarded as a significant threat to the production of safe food and the arise of antibiotic-resistant bacteria. The objective of this investigation is to evaluate the quorum sensing inhibitory effect of Nepeta curviflora methanolic extract. Methods: The effectiveness of the leaves at sub-inhibitory concentrations of 2.5, 1.25, and 0.6 mg/mL on the virulence factors and biofilm formation of P. aeruginosa was evaluated. The effect of N. curviflora methanolic extract on the virulence factors of P. aeruginosa, including pyocyanin, rhamnolipid, protease, and chitinase, was evaluated. Other tests including the crystal violet assay, scanning electron microscopy (SEM), swarming motility, aggregation ability, hydrophobicity and exopolysaccharide production were conducted to assess the effect of the extract on the formation of biofilm. Insight into the mode of antiquorum sensing action was evaluated by examining the effect of the extract on the activity of N-Acyl homoserine lactone (AHL) and the expression of pslA and pelA genes. Results: The results showed a significant attenuation in the production of pyocyanin and rhamnolipid and in the activities of protease and chitinase enzymes at 2.5 and 1.25 mg/mL. In addition, N. curviflora methanolic extract significantly inhibited the formation of P. aeruginosa biofilm by decreasing aggregation, hydrophobicity, and swarming motility as well as the production of exopolysaccharide (EPS). A significant reduction in AHL secretion and pslA gene expression was observed, indicating that the extract inhibited quorum sensing by disrupting the quorum-sensing systems. The quorum-sensing inhibitory effect of N. curviflora extract appears to be attributed to the presence of kaempferol, quercetin, salicylic acid, rutin, and rosmarinic acid, as indicated by LCMS analysis. Conclusion: The results of the present study provide insight into the potential of developing anti-quorum sensing agents using the extract and the identified compounds to treat infections resulting from quorum sensing-mediated bacterial pathogenesis.

Characterization of Extended Spectrum Beta-Lactamases (ESBL) Producing Escherichia coli Isolates from Surface Water Adjacent to Pharmaceutical Industries in Bangladesh: Antimicrobial Resistance and Virulence Pattern

  • Taslin Jahan Mou;Nasrin Akter Nupur;Farhana Haque;Md Fokhrul Islam;Md. Shahedur Rahman;Md. Amdadul Huq;Anowar Khasru Parvez
    • Microbiology and Biotechnology Letters
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    • v.51 no.3
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    • pp.268-279
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    • 2023
  • The pharmaceutical industry in Bangladesh produces a diverse range of antibiotics for human and animal use, however, waste disposal management is inadequate. This results in substantial quantities of antibiotics being discharged into water bodies, which provide suitable environment for the growth of antibiotic-resistant bacteria, capable of spreading resistance genes. This study intended for exploring the bacterial antibiotic resistance profile in adjoining aquatic environmental sources of pharmaceutical manufacturing facilities in Bangladesh. Seven surface water samples were collected from the vicinity of two pharmaceutical industries located in the Savar area and 51 Escherichia coli isolates were identified using both phenotypic and genotypic methods. Antibiotic susceptibility tests revealed the highest percentage of resistance against ampicillin, azithromycin, and nalidixic acid (100%) and the lowest resistance against meropenem (1.96%) out of sixteen different antibiotics tested. 100% of the study E. coli isolates were observed with Multidrug resistance phenotypes, with the Multiple Antibiotic Resistance (MAR) value ranging from 0.6-1.0. Furthermore, 69% of the isolates were Extended Spectrum Beta-Lactamases (ESBL) positive as per the Double Disk Diffusion Synergy Test (DDST). ESBL resistance genes blaTEM, blaCTX-M-13, blaCTX-M-15, and blaSHV were detected in 70.6% (n = 36), 60.8% (n = 32), 54.9% (n = 28), and 1.96% (n = 1) of the isolates, respectively, by Polymerase Chain Reaction (PCR). Additionally, 15.68% (n = 8) of the isolates were positive for E. coli specific virulence genes in PCR. These findings suggest that pharmaceutical wastewater, if not properly treated, could be a formidable source of antibiotic resistance spread in the surrounding aquatic environment. Therefore, continued surveillance for drug resistance among bacterial populations around drug manufacturing facilities in Bangladesh is necessary, along with proper waste disposal management.

Antioxidant Activity of Native Korean Halophyte Extracts and Their Anti-biofilmActivity against Acinetobacter baumannii (한국 자생 염생식물 추출물의 항산화 활성 및 다재내성 Acinetobacter baumannii에 대한 항생물막 활성)

  • Eun Seong Lee;Jeong Woo Park;Ki Hwan Moon;Youngwan Seo
    • Journal of Life Science
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    • v.33 no.12
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    • pp.1015-1024
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    • 2023
  • Antibiotics have greatly contributed to the treatment and prevention of bacterial diseases in humans, animals, and fish. However, antibiotic misuse has led to the emergence and spread of multidrug-resistant bacteria. In addition to antibiotic discovery research, efforts are being made to combat such multidrug-resistant bacteria using antimicrobial agents, antioxidants, host immune enhancement, probiotics, and bacteriophages, as well as various symptomatic therapies. To discover novel bioactive compounds, it is crucial to adopt approaches that incorporate fresh ideas, new targets, innovative techniques, and untapped resources. Halophytes are plants that grow in high-salt soils and are known to adapt to salt-induced stress through unique metabolic processes that produce secondary metabolites. This study aimed to investigate the effects of extracts of halophytes native to Korea on oxidative stress and to determine whether they exert inhibitory activity against biofilms, which are major pathogenic factors of infectious bacteria. The Acinetobacter baumannii strain ATCC 17978, a representative drug-resistant bacterium, was used to measure anti-biofilm activity. The results showed that Aster spathulifolius, Carex kobomugi, Rosa rugosa, and Asparagus cochinchiensis exerted strong antioxidant and anti-biofilm effects without affecting bacterial growth itself. The halophytes used in this study are promising candidates for the development of pharmaceutical agents with antioxidant and antimicrobial properties.

Bacterial Meningitis in Children in One Tertiary Hospital (소아의 세균성 수막염)

  • Oh, Ji Eun;Chang, Ji Yeon;Kwon, Young Se;Kim, Soon Ki;Son, Byong Kwan;Hong, Young Jin
    • Pediatric Infection and Vaccine
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    • v.10 no.2
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    • pp.208-214
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    • 2003
  • Purpose : We performed a retrospective study on bacterial meningitis in children, pertaining to the causative organisms, bacterial resistance to antibiotics and the effect of recent introduction of Haemophilus influenzae type b(Hib) vaccine. Methods : We analysed the forty-three cases of bacterial meningitis which had been treated at the Inha University Hospital from June 1996 to June 2003. Results : Nineteen cases(44.2%) of them were infants younger than 2 months of age, and 29 cases(67.5%) younger than 1 year of age. The common causative organisms under 2 months of age were group B streptococcus(GBS)(47.4%), E. coli(21.1%), and Klebsiella pneumoniae(21.1%). In the age group beyond 2 months of age, S. pneumoniae were seen in 50 %, H. influenzae in 16.7% and N. meningitidis in 16.7%. All of the five cases of Hib meningitis had not been vaccinated for Hib. There has been no Hib meningitis cases since 2001. Overall fatality rate was 4.5%, and complication occurred in 39%. Complications was significantly less frequent in patients resumed to be treated within 48 hours after onset compared to after 48 hours after onset. Penicillin resistance of S. pneumoniae and GBS isolated among bacterial meningitis cases was high. Conclusion : Timing of treatment after the onset of the disease appeared the most important factor for prognosis of bacterial meningitis. The cases of H. influenzae meningitis have decreased probably due to Hib vaccination.

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Treatment test for bovine mastitis by the determination of ATP based on firefly bioluminescence (Bioluminescence 반응에 의한 ATP측정을 이용한 젖소 유방염 치료에 관한 연구)

  • Kim, Tae-jong;Kim, Jong-bae;Lee, Seong-bae;Jeon, Young-soo
    • Korean Journal of Veterinary Research
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    • v.29 no.3
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    • pp.393-405
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    • 1989
  • This study was carried out to treatment test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are followed; 1. In the susceptibility test, cephalothin which looks the most effective were sensitive to Staphylococcus sp. (72.3%), Micrococcus sp. (84.2%), Streptococcus sp. (72.7%) and Gram positive bacilli (72.7%), Gram negative bacilli were sensitive to gentamicin (92.3%) and Yeast-like-fungi was the most sensitive to clotrimazole, and nystatin in order. 2. When the number of bacteria, such as Staphylococcus aureus, Candida tropicalis isolated from the mastitis milk were counted by conventional agar plating technique, and compared with the concentration of bacterial ATP, it gave a good linear relationship. The content of ATP per Staphylococcus aureus, cell was 3.1fM and Candida tropicalis showed the high level of A TP (90fM). 3. The ATP assay was applied to the determination of minimal inhibitory concentration (MIC) of various antibiotics. When Staphylococcus aureus was incubated in the presence of different concentration of tetracycline, erythromycin, kanamycin and streptomycin sulfate and the growth was monitored by the conventional agar plating technique and ATP assay, both methods shown the same results that they were 1mcg/ml, 2mcg/ml, 6.25mcg/ml and 8mcg/ml, respectively. 4. For the determination of susceptibility of sensitive and resistant Staphylococcus au reus isolated for the milk with mastitis to tetracycline, erythromycin kanamycin and streptomycin sulfate, the minimum time required for the test was determined by the assay of ATP every 30 minutes during incubation of 3 hours at $37^{\circ}C$. ATP concentration time curve calculated on both resistant and sensitive strains incubated 3 hours as the optimum time for the determination of susceptibilities of various antibiotics exemed. The ATP concentration of each test broth (antibiotic containing), expressed as a percentage of its own control brith (antibioticfree) indicated values of 30% to be indicative of each antibiotic sensitivity. Single time point ATP assay carried out on the various sensitive and resistant of Staphylococcus aureus to antibiotics examined after 3 hours at $37^{\circ}C$ correlated exactly with disc diffusion and MIC. 5. In the cure of intramammary treatment of bovine mastitis in lactating quarters, the cure rate of Staphylococcal mastitis showed to cephalexin (80%), cloxacillin and gentamicin (70%), ampicillin and oxytetracycline (60%), and Streptococcal mastitis showed to cephalexin (85%), penicillin (80%), cloxacillin and oxytetracycline (75%), and ampicillin (70%), but intramammary antimycotic drug (clotrimazol) were only a little effect about fungal mastitis.

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The utilization of antibiotics and the treatment of bacterial diseases in fish (항생제 사용과 세균성 어류질병의 치료)

  • Jeong, Hyun-Do;Chun, Seh-Kyu
    • Journal of fish pathology
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    • v.5 no.1
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    • pp.37-48
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    • 1992
  • Antibiotics are chemical substances produced by various species of microorganisms that suppress the growth of other microorganisms or may destroy them. Among the more than 4000 antibiotics that has been identified, about 20s are using as the therapy of infectious fish diseases. There are several methods used to classify and group antibiotics, and the most common classification has been based on chemical structure and proposed mechanism of action. The effect of antibiotics may be determined by the kind of fish pathogens and by the external environment surrounded the infected fish. It implies that the kind of antibiotics and its application method should be decided after the determination of the reasons of fish disease. The uncontrolled usages of antibiotics may induce the selection of resistant mutants appeared spontaneously and present in any group of bacteria. The epidemic spread of such antibiotic resistant strains of fish pathogenic bacteria already has been reported in various districts of japan. Importantly, transferable drug resistant(R) plasmids were detected in strains of most of fish pathogens. Based on those reports, the antimicrobial resistance appears to be a rapidly emerging problem in the fish industry on the country. The expanding literatures on the pharmacokinetics, clinical trials, withdrawal periods and efficacy of environmental effect for the commonly using antibiotics have met the needs of data for the practical application of antibiotics. However, the most important thing for the treatment of fish diseases would be the communication and exchanging of information between the site of aquaculture and the diagnostic laboratory.

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Characterization of antimicrobial proteins produced by Bacillus sp. N32 (Bacillus sp. N32 균주가 생산하는 항균 단백질 특성)

  • Lee, Mi-Hye;Park, In-Cheol;Yeo, Yun-Soo;Kim, Soo-Jin;Yoon, Sang-Hong;Lee, Suk-Chan;Chung, Tae-Young;Koo, Bon-Sung
    • The Korean Journal of Pesticide Science
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    • v.10 no.1
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    • pp.56-65
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    • 2006
  • An antagonistic bacterial isolate, that inhibits the growth of plant pathogens, was selected and identified from 5,000 isolates screened from the rhizosphere of various crop plants. An isolate Bacillus sp. N32, tested against Colletotrichum gloeosporioides causing anthracnose disease in hot pepper, produced both a heat resistant antifungal protein and a heat sensitive antifungal protein. The heat resistant protein was partially purified by Ammonium sulfate fractionation and gel filtration chromatography. The bioautography showed that the proteins possessed high antifungal activity. The biosynthetic gene cluster responsible for the heat resistant antifungal protein was cloned from cosmid library using DNA probe obtained from PCR product with the primers targeting the conserved nucleotide sequence of the synthetic genes reported earlier, Most of the clones obtained showed higher homology to fengycin antibiotic synthetic gene family reported earlier. On the other hand, the heat sensitive protein was isolated from SDS-PAGE and electroblotting to determine the N-terminal amino acid sequences. The heat sensitive antifungal protein gene was cloned from the ${\lambda}-ZAP$ libraries using a DNA probe based on the N-terminal amino acid sequences of the heat sensitive protein. We are contemplating to clone and sequence the whole gene cluster encoding the heat sensitive protein for further analysis.

Community-acquired Extended-spectrum and Plasmid-mediated ampC Beta-lactamase-producing Multidrug-resistant Enterobacter cloacae Septicaemia in a Cat with Euthyroid Sick Syndrome (정상 갑상샘 질환 증후군 고양이의 지역사회획득 광범위 및 플라스미드 유래 ampC beta-lactamase 양성 다약제내성 Enterobacter cloacae 패혈증)

  • Han, Jae-Ik;Na, Ki-Jeong
    • Journal of Veterinary Clinics
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    • v.32 no.2
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    • pp.191-195
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    • 2015
  • A 7-year-old castrated male Korean Shorthair cat was referred with lethargy and anorexia. Laboratory examination revealed moderate degenerative changes of peripheral neutrophils on blood smear examination and decreased levels of free and total thyroxine ($T_4$) as well as bacterial growth on blood culture. Molecular analyses of the 16S ribosomal RNA gene and heat shock protein 60 gene confirmed the bacterium as Enterobacter cloacae. A minimal inhibitory concentration test showed multidrug resistance of the bacterium against 16 antibiotics. Polymerase chain reaction (PCR) and subsequent sequencing specifically for $bla_{TEM}$, $bla_{SHV}$, $bla_{CTX-M}$, and plasmid-mediated ampC genes revealed positive results to $bla_{TEM-1}$, $bla_{CTX-M-15}$, and plasmid-mediated $bla_{ACT-1}$ genes, indicating that the isolated bacterium contains plasmids containing genes encoding extended-spectrum beta-lactamase and plasmid-mediated ampC beta-lactamase. After 1 month of treatment with antibiotics and levothyroxine, the cat's condition improved; both the thyroid function test and the blood culture showed no abnormalities. This is the first report of community-acquired multidrug-resistant E. cloacae-induced euthyroid sick syndrome in a cat. By the prompt diagnostic procedures and properly selected antibiotic therapy, the cat was recovered from the multidrug-resistant bacterium-induced septicaemia.

Antibiotic sensitivity of the bacterial strains isolated from operating wounds (수술창에서 분리된 균주의 항생제 감수성)

  • Oh, Yang-Hyo;Kim, Yung-Bu;Park, Young-Min;Kim, Min-Jung;Cha, Mi-Sun
    • Journal of Life Science
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    • v.8 no.1
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    • pp.40-50
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    • 1998
  • Staphylococuus aureus and Escherichia coli is increasingly responsible for outbreaks of nosocomial infection around the world. Because serious infections due to these organisms currently necessiate use of non-$\beta$-lactam antimicrobial therapy and because strains is ofen resistant to many antimicrobial agents, infections with this organism are difficult to treat. Isolated strains from post operaton wounds of PNU hospital patient were tested for the antimicrobial susceptibility, resistant pattern and combined action to the 6 antibiotics. The minimal inhibitory concentraction of each antibiotic anc antibiotics combining in various ratios were measured by checkerboard dilution method. the synergism was determined through calculating the fractional inhibitory concentraction index (FICI). In case of S. aureus, 15 strains was shown to be highly sensitive to streptomycin and 13 strains to cephalothin. In case of E. coli, it is excellent senstitive 16 strains, sensitive 4 strains on cefoperazone, as like S. aureus, and thus the sensitive is most to be 66%. As the result of gaining MIC from S. aureus upon agar dilution method, MIC$_{50}$ was 8$\mu$g/ml, MIC$_{90}$ was 16$\mu$g/ml and thus the streptomycine is shown to be lowest. In case of E, coli, S. MIC$_{50}$ was 4$\mu$g/ml, MIC$_{90}$ was 16$\mu$g/ml, in streptomycin and thus is shown to be lower than S. aureus. As the result of comparing the resistance aspect of combining the antibiotics on S. aureus and E. coli, the resistant strain can be known to be reduced to the large range more than each 40% than combining with only aminoglycoside-series or cephalosporine-series. As the result of combining aminoglycoside-series, streptomycin and cephalothin or cefuroxime sensitive to S. aureus and E. coli in the above mentioned results, the increase or imporovement of effect is over 73% and 80%, respectively, thus the case od combining 2 antibiotics is shown to be better in the effect. Isolated strains from operating wounds were for the antimicrobial susceptibility. In case of S. aureus 15 strains was shown to be sensitive very much on streptomycin. In case of E. coli it is excellent sensitive 16 strains. As the results of combining aminoglycosides-series, streptomycin and cephalosporine series, cephalothin and cefuroxime, the increase or improvement of effect is over 73%, thus case of combining 2 antibiotics is shown to be better in the effect.

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