• Title/Summary/Keyword: isolation, identification

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Secondary Metabolites from Anthonotha cladantha (Harms) J.Léonard

  • Laurent Voufack Lefack Bongmo;Achille Nouga Bissoue;Samuel Magloire Bissim;Georges Bellier Tabekoueng;Willifred Dongmo Tekapi Tsopgni;Mehreen Lateef;Felicien Mushagalusa Kasali;Muhammad Shaiq Ali;Alain Francois Kamdem Waffo;Jean Duplex Wansi
    • Natural Product Sciences
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    • v.29 no.1
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    • pp.50-58
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    • 2023
  • The phytochemical investigation of the crude methanolic extracts roots and stem bark of Anthonotha cladantha (Harms) J.Léonard led to the isolation and identification of twelve secondary metabolites: 2,3-dihydroxypropyl hexacosanoate (1), hederagenine (2), cycloeucalenol (3), 2α-hydroxylupeol (4), betulinic acid (5), lupeol (6), heptacosan-2-one (7), triacontanoic acid (8), stigmast-4-en-3-one (9), β-sitosterol (10), stigmasterol (11), and stigmasterol-3-O-β-D-glucopyranoside (12). Their structures were elucidated with the help of their spectroscopic and physical data and by comparison with those reported in the literature. To the best of our knowledge, from all those compounds, 2,3-dihydroxypropyl hexacosanoate (1), hederagenine (2), cycloeucalenol (3), 2α-hydroxylupeol (4), and betulinic acid (5) are being reported for the first time from this genus. In addition, the acetylation of compound 1 afforded a new derivative 3-(hexacosanoyloxy)propane-1,2-diyl diacetate (1a). Compound 1 possessed a moderate α-glucosidase inhibitory activity with an IC50 value of 39.2 ± 0.22 μM; it neither showed antioxidant activity nor inhibition against the enzyme urease. Compound 1a exhibited weak antioxidant activity in the DPPH assay with an IC50 value of 80.3 ± 0.83 μM but was inactive against α-glucosidase and urease. Furthermore, both compounds 1 and 1a were inactive against seven pathogenic bacterial strains.

Isolation and identification of mosquito-borne zoonotic diseases in slaughterhouse in Daejeon

  • Youngju Kim;Gyurae Kim;Sunkyong Song;Youngshik Jung;Seojin Park;Sang-Joon Lee;Ho-Seong Cho;Yeonsu Oh
    • Korean Journal of Veterinary Service
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    • v.46 no.2
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    • pp.115-122
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    • 2023
  • This study was performed to investigate the distribution of mosquito vectors related to the zoonotic disease in Daejeon. Samples were taken using a blacklight trap once a month from March to November 2021 at the slaughterhouse in Daejeon. A total of 820 mosquitoes were captured and classified into 5 genera and 8 species. Among the collected mosquitoes, 319 (38.9%) and 295 (35.93%) were Aedes vexans nipponii and Culex pipiens pallens, respectively, making them the dominant species. The overall number of mosquitoes collected started to increase from May and reached the largest value of 329 (40.12%) in June. Trapped mosquitoes are created 72 pools by environmental condition and by species. The pools were tested by PCR methods for 7 zoonotic pathogens. Flavivirus-positive products were confirmed by DNA sequencing. Japanese encephalitis viruses were detected in 3 pools collected from cow lairage (Culex pipiens pallens) in May, cow by-product processing room (Aedes vexans nipponii) in June and cow lairage (Mansonia uniformis) in June. Culex flavivirus were detected in 4 pools. Based on the results of this study, it is considered that continous surveillence of mosquitoes in livestock assembly facilities (slaughterhouse) should be performed for controlling mosquito populations and mediating disease spread by mosquitoes.

Genomic Analysis of the Carrot Bacterial Blight Pathogen Xanthomonas hortorum pv. carotae in Korea

  • 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.

Characterization of degradation products of the Balsalazide by Mass spectrometry: Optimization of stability-indicating HPLC method for separation and quantification of process related impurities of Balsalazide

  • Chilakabattina Naga Narasimha Babu;Ch. Srinivasa Reddy;Bhagya Kumar Tatavarti;M. Radha Madhavi;Venkateswara Rao Anna
    • Analytical Science and Technology
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    • v.37 no.1
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    • pp.25-38
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    • 2024
  • The study aimed to investigate a novel approach by utilizing liquid chromatography (LC) and liquid chromatography-mass spectrometry (LC-MS) to separate, identify and characterize very nominal quantities of degradation products (DPs) of balsalazide along with its process related impurities without isolation from their reaction mixtures. The impurities along with balsalazide were resolved on spherisorb ODS2 (250×4.6 mm, 5.0 ㎛) column at room temperature using 0.2 M sodium acetate solution at pH 4.5 and methanol in the ratio of 55:45 (v/v) as mobile phase pumped isocratically at 1.0 mL/min as mobile phase and UV detection at 255 nm. The method shows sensitive detection limit of 0.003 ㎍/mL, 0.015 ㎍/mL and 0.009 ㎍/mL respectively for impurity 1, 2 and 3 with calibration curve liner in the range of 50-300 ㎍/mL for balsalazide and 0.05-0.30 for its impurities. The balsalazide pure compound was subjected to stress studies and a total of four degradation products (DPs) were formed during the stress study and all the DPs were characterized with the help of their fragmentation pattern and the masses obtained upon LC-MS/MS. The DPs were identified as 3-({4-[(E)-(4-hydroxyphenyl) diazenyl]benzoyl}amino)propanoic acid (DP 1), 4-[(E)-(4-hydroxyphenyl)diazenyl] benzamide (DP 2), 5-[(E)-(4-carbamoylphenyl)diazenyl]-2-hydroxybenzoic acid (DP 3) and 3-({4-[(E)-phenyldiazenyl]benzoyl}amino)propanoic acid (DP 4). Based on findings, it was concluded that, the proposed method was successfully applicable for routine analysis of balsalazide and its process related impurities in pure drug and formulations and also applicable for identification of known and unknown impurities of balsalazide.

Administrative codes may have limited utility in diagnosing biliary colic in emergency department visits: A validation study

  • Jordan Nantais;Muhammad Mansour;Charles de Mestral;Shiva Jayaraman;David Gomez
    • Annals of Hepato-Biliary-Pancreatic Surgery
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    • v.26 no.3
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    • pp.277-280
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    • 2022
  • Backgrounds/Aims: Biliary colic is a common cause of emergency department (ED) visits; however, the natural history of the disease and thus the indications for urgent or scheduled surgery remain unclear. Limitations of previous attempts to elucidate this natural history at a population level are based on the reliance on the identification of biliary colic via administrative codes in isolation. The purpose of our study was to validate the use of International Statistical Classification of Diseases and Related Health Problems codes, 10th Revision, Canadian modification (ICD-10-CA) from ED visits in adequately differentiating patients with biliary colic from those with other biliary diagnoses such as cholecystitis or common bile duct stones. Methods: We performed a retrospective validation study using administrative data from two large academic hospitals in Toronto. We assessed all the patients presenting to the ED between January 1, 2012 and December 31, 2018, assigned ICD-10-CA codes in keeping with uncomplicated biliary colic. The codes were compared to the individually abstracted charts to assess diagnostic agreement. Results: Among the 991 patient charts abstracted, 26.5% were misclassified, corresponding to a positive predictive value of 73% (95% confidence interval 73%-74%). The most frequent reasons for inaccurate diagnoses were a lack of gallstones (49.8%) and acute cholecystitis (27.8%). Conclusions: Our findings suggest that the use of ICD-10 codes as the sole means of identifying biliary colic to the exclusion of other biliary pathologies is prone to moderate inaccuracy. Previous investigations of biliary colic utilizing administrative codes for diagnosis may therefore be prone to unforeseen bias.

Isolation, Identification and Use of Bacterial Strain Ochrobactrum intermedium PDB-3 for Degradation of the Pesticide Chlorpyrifos

  • Diyorbek Kosimov;Lyudmila Zaynitdinova;Aziza Mavjudova;Muzaffar Muminov;Oybek Shukurov
    • Microbiology and Biotechnology Letters
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    • v.52 no.1
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    • pp.44-54
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    • 2024
  • One of the serious modern environmental problems is pollution caused by highly toxic pesticides. Only small amounts of applied pesticides reach their target, and the rest ends up in soil and water. Chlorpyrifos is a toxic, broad-spectrum organophosphate insecticide. In humans, chlorpyrifos inhibits acetylcholinesterase (AChE) in the peripheral and central nervous system, and particularly in children, small amounts of this pesticide cause neurotoxic damage. As the toxic effects of chlorpyrifos and its persistence in the environment require its removal from contaminated sites, it is essential to study the biological diversity of chlorpyrifos-degrading microorganisms. In this study, we sought to determine the chlorpyrifos-degrading ability of the bacterial strain Ochrobactrum intermedium PDB-3. This strain was isolated from soil contaminated with various pesticides and identified as PDB-3 based on morpho-cultural characteristics, MALDI-TOF MS, and 16S rRNA. Studies were conducted for 30 days in sterile soils containing initial concentrations of 50, 75, 100, and 125 mg/kg of chlorpyrifos. To determine the degradation of chlorpyrifos, a liquid culture of the strain was added to the soil at three optical densities: 0, and after 24 and 48 h (OD = 0.03, 0.2 and 0.32). Using GX-MS, we determined that chlorpyrifos was converted to 3,5,6-trichloro-2-pyridinol (TCP). We also found that with increasing optical density, rapid degradation of the initial concentration of chlorpyrifos occurred. Sterile soil without strain PDB-3 was used as a control sample.

A Novel Draft Genome-Scale Reconstruction Model of Isochrysis sp: Exploring Metabolic Pathways for Sustainable Aquaculture Innovations

  • Abhishek Sengupta;Tushar Gupta;Aman Chakraborty;Sudeepti Kulshrestha;Ritu Redhu;Raya Bhattacharjya;Archana Tiwari;Priyanka Narad
    • Microbiology and Biotechnology Letters
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    • v.52 no.2
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    • pp.141-151
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    • 2024
  • Isochrysis sp. is a sea microalga that has become a species of interest because of the extreme lipid content and rapid growth rate of this organism indicating its potential for efficient biofuel production. Using genome sequencing/genome-scale modeling for the prediction of Isochrysis sp. metabolic utilities there is high scope for the identification of essential pathways for the extraction of byproducts of interest at a higher rate. In our work, we design and present iIsochr964, a genome-scale metabolic model of Isochrysis sp. including 4315 reactions, 934 genes, and 1879 metabolites, which are distributed among fourteen compartments. For model validation, experimental culture, and isolation of Isochrysis sp. were performed and biomass values were used for validation of the genome-scale model. OptFlux was instrumental in uncovering several novel metabolites that influence the organism's metabolism by increasing the flux of interacting metabolites, such as Malonyl-CoA, EPA, Protein and others. iIsochr964 provides a compelling resource of metabolic understanding to revolutionize its industrial applications, thereby fostering sustainable development and allowing estimations and simulations of the organism metabolism under varying physiological, chemical, and genetic conditions. It is also useful in principle to provide a systemic view of Isochrysis sp. metabolism, efficiently guiding research and granting context to omics data.

Microbial and Pathogenic Contamination of Ready-to-eat Fresh Vegetables in Korea (한국에 유통중인 신선편이 채소류의 미생물 품질 및 병원성 세균의 오염도 조사)

  • Bae, Young-Min;Hong, Yu-Jin;Kang, Dong-Hyun;Heu, Sung-Gi;Lee, Sun-Young
    • Korean Journal of Food Science and Technology
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    • v.43 no.2
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    • pp.161-168
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    • 2011
  • The purpose of this study was to evaluate microbiological contamination of fresh vegetables in Korea. Twenty types of vegetables were tested for total aerobic bacteria, coliforms, Escherichia coli, yeast and mold, and pathogenic bacteria such as Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Salmonella, E. coli O157:H7, Cronobacter sakazakii, Shigella, and Campylobacter. Levels of total aerobic bacteria and coliform on 20 vegetables were between 3.74 and 8.04 log CFU/g, and 0.16 and 5.02 log CFU/g, respectively. The highest contamination levels of total aerobic bacteria were observed on water dropwort, sprouts, mungbean sprout, and ballflower root. There was no significant difference (p>0.05) in microbial contamination levels of total aerobic count, coliform, E. coli, yeast and mold between organic and nonorganic vegetables. When isolation methods using selective agars were applied, L. monocytogenes, B. cereus, Salmonella and Campylobacter were isolated from some fresh vegetable samples. Results of API kit tests showed that L. monocytogenes was identified on Chinese cabbage, cucumber, soybean sprouts, and iceberg lettuce while Salmonella was identified on Korean leek. Furthermore, Campylobacter jejuni was also identified in more than 50 of the 100 samples. However, when positive samples from API kit were tested for real-time PCR or 16S rRNA sequencing method, only B. cereus from perilla leaf, carrot, water dropwort, and sprouts showed positive results. These results indicate that selective agar and API kit detection methods might result in false positive results for some pathogens. Therefore, studies need to improve isolation or confirmation methods for such pathogens.

Isolation and Morphological Identification of Fresh Water Green Algae from Organic Farming Habitats in Korea (유기농업 생태계로부터 담수 녹조류 분리 및 형태적 동정)

  • 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
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    • 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.

Screening and Identification of a Cesium-tolerant Strain of Bacteria for Cesium Biosorption (환경유래의 세슘 저항성 균주 선별 및 세슘 흡착제거 연구)

  • Kim, Gi Yong;Jang, Sung-Chan;Song, Young Ho;Lee, Chang-Soo;Huh, Yun Suk;Roh, Changhyun
    • Korean Journal of Environmental Biology
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    • v.34 no.4
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    • pp.304-313
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    • 2016
  • One of the issues currently facing nuclear power plants is how to store spent nuclear waste materials which are contaminated with radionuclides such as $^{134}Cs$, $^{135}Cs$, and $^{137}Cs$. Bioremediation processes may offer a potent method of cleaning up radioactive cesium. However, there have only been limited reports on $Cs^+$ tolerant bacteria. In this study, we report the isolation and identification of $Cs^+$ tolerant bacteria in environmental soil and sediment. The resistant $Cs^+$ isolates were screened from enrichment cultures in R2A medium supplemented with 100 mM CsCl for 72 h, followed by microbial community analysis based on sequencing analysis from 16S rRNA gene clone libraries(NCBI's BlastN). The dominant Bacillus anthracis Roh-1 and B. cereus Roh-2 were successfully isolated from the cesium enrichment culture. Importantly, B. cereus Roh-2 is resistant to 30% more $Cs^+$ than is B. anthracis Roh-1 when treated with 50 mM CsCl. Growth experiments clearly demonstrated that the isolate had a higher tolerance to $Cs^+$. In addition, we investigated the adsorption of $0.2mg\;L^{-1}$ $Cs^+$ using B. anthracis Roh-1. The maximum $Cs^+$ biosorption capacity of B. anthracis Roh-1 was $2.01mg\;g^{-1}$ at pH 10. Thus, we show that $Cs^+$ tolerant bacterial isolates could be used for bioremediation of contaminated environments.