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Bacterial Population Dynamics of Natural Fermentation of Sumbawa Mare’s Milk Using Metagenomic Approach 用宏基因组法研究松巴哇马奶自然发酵细菌种群动态
Microbiology Indonesia Pub Date : 2021-06-28 DOI: 10.5454/mi.15.2.2
Y. Jatmiko, A. Suharto, I. Mustafa, Siska Aditya
{"title":"Bacterial Population Dynamics of Natural Fermentation of Sumbawa Mare’s Milk Using Metagenomic Approach","authors":"Y. Jatmiko, A. Suharto, I. Mustafa, Siska Aditya","doi":"10.5454/mi.15.2.2","DOIUrl":"https://doi.org/10.5454/mi.15.2.2","url":null,"abstract":"This study aimed to assess the changing of bacterial density and the physicochemical aspects during natural fermentation of Sumbawa mare’s milk, and to evaluate the dynamics of bacterial population during the natural fermentation using metagenomic approach. Mare’s milk sample obtained from Regency of Dompu were fermented for 60 days. On the day 0, 7, 15, 30 and 60 mare milk sample were collected for further analysis, such as bacterial density enumeration, nutrition content, physical properties of the milk, and total DNA isolation. The total DNA samples obtained were analyzed using next generation sequencing. The density of lactic acid bacteria was decreased along with fermentation periods. Meanwhile, the density of aerobic bacteria on was relatively fluctuated. The physicochemical content of mare’s milk also changed during fermentation periods. Carbohydrate content and total sugar was decrease along with the decreasing of pH value. Moreover, the lipid content increase, and the protein content was fluctuated. The changing in physical properties such as whey color, acidity and gas was observed until the end of mare’s milk natural fermentation process. Using metagenomics analysis, the bacterial diversity from each sample periods categorized as low because of the dominance of Lactobacillus helveticus until the end of the fermentation. Lactobacillus helveticus as a member of LAB did not grow on isolation media on the late stage of fermentation periods (day-60). The presence of uncultivable bacteria can be detected with metagenomic approach, fulfilling the limited information on the bacterial composition of fermented Sumbawa mare’s milk products.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86155536","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
Thermostable Alkaline Protease Activity from Aspergillus flavus DUCC- K225 and Its Compatibility to Local Detergents 黄曲霉DUCC- K225耐热碱性蛋白酶活性及其与局部洗涤剂的相容性
Microbiology Indonesia Pub Date : 2021-04-08 DOI: 10.5454/MI.15.1.3
A. Rini, Isworo Rukmini, S. Pujiyanto
{"title":"Thermostable Alkaline Protease Activity from Aspergillus flavus DUCC- K225 and Its Compatibility to Local Detergents","authors":"A. Rini, Isworo Rukmini, S. Pujiyanto","doi":"10.5454/MI.15.1.3","DOIUrl":"https://doi.org/10.5454/MI.15.1.3","url":null,"abstract":"Protease is the most important enzyme in many industries, including detergent. Aspergillus flavus DUCC-K225 is a thermotolerant indigenous molds isolated from Madura island which is potential in producing thermostable alkaline protease enzymes. The enzyme produced by submerged culture on modified Czapeks Dox liquid medium containing glucose as carbon source and 1% of casein. The aims of this study were to determine the activity and stability of thermostable alkaline protease produced by A. flavus DUCC-K225 at various temperatures, also the compatibility to 5 local detergents. Research were done using CRD, with temperature variation for protease activity as treatment. The results showed that the optimum activity of thermostable alkaline proteases was 214.503 U/mL, with retained activities up to 78% in 60 minutes at 55°C.  The alkaline thermostable protease compatible with 5 local detergents tested, with the retained activity varied 55.8%-95.22% at 29oC and 62.36%-97.7% at 55oC. The compatibility to detergent confirmed by blood stained cleansed. The higher the protease activity, the higher the blood stain solubility. \u0000  \u0000Key words: Alkaline protease, thermostable, A. flavus DUCC-K225, compatibility, detergent","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90874307","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Growth Kinetic Study of Blue-green Microalgae Arthrospira platensis Using Buffalo Manure as Alternative Media 水牛粪替代蓝绿微藻平台节螺旋藻生长动力学研究
Microbiology Indonesia Pub Date : 2021-03-31 DOI: 10.5454/MI.14.4.5
M. M. Martgrita, Brian Sinaga, Lianty Simangunsong, Andy Trirakhmadi, M. Pasaribu
{"title":"Growth Kinetic Study of Blue-green Microalgae Arthrospira platensis Using Buffalo Manure as Alternative Media","authors":"M. M. Martgrita, Brian Sinaga, Lianty Simangunsong, Andy Trirakhmadi, M. Pasaribu","doi":"10.5454/MI.14.4.5","DOIUrl":"https://doi.org/10.5454/MI.14.4.5","url":null,"abstract":"North Sumatera is one of the provinces in Indonesia with the highest buffalo population, which is responsible for the high accumulation of buffalo manure that can cause environmental and aesthetic problems if left untreated. One of the possible alternatives for solving this issue is by implementing buffalo manure as growth media for microorganisms, e.g. microalgae. In this research, buffalo manure was used as alternative media for Arthospira platensis cultivation. Buffalo manure was taken from Sitoluama village, Laguboti, Toba Regency of North Sumatra Province. Research steps included media and culture preparation, cultivation, sampling, sample analysis and verification of constructed models and validation. Buffalo manure concentration in media is varied from 1 g.L-1 to 8 g.L-1 which is analogous to nitrogen content of 0.002 mg.L-1 to 0.018 mg.L-1. Growth data was used for growth kinetic modelling, which was most satisfactory for Monod model (µmax = 0.5915 day-1, Ks = 0.421 g.L-1).","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83624067","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Trench Construction in Peat Soil and the DGGE Analyses of Nif Gene and Activity of Dehydrogenase 泥炭沟渠建设及Nif基因和脱氢酶活性的DGGE分析
Microbiology Indonesia Pub Date : 2021-03-12 DOI: 10.5454/MI.14.4.2
H. Widiastuti, D. Taniwiryono, Iman Rusmana, G. Permatasari
{"title":"Trench Construction in Peat Soil and the DGGE Analyses of Nif Gene and Activity of Dehydrogenase","authors":"H. Widiastuti, D. Taniwiryono, Iman Rusmana, G. Permatasari","doi":"10.5454/MI.14.4.2","DOIUrl":"https://doi.org/10.5454/MI.14.4.2","url":null,"abstract":"Basal stem rot (BSR) is a major disease in oil palm crops which also possible happen on peat soils. Ganoderma boninense is the pathogen that causes this disease infects the plants through the root. Maintaining root health expects to reduce the intensity of the occurrence of BSR disease and its spread. In this research, trenching construction and maintenance is carried out which aims to break the spread and activity of Ganoderma. Trenching maintenance is carried out by giving empty bunches oil palm inoculated with decomposer, Azotobacter inoculants and humic acid. Four treatments tested were trenched (T1) and untrenched (T0) in combination with two level of Ganoderma attack, which are moderate (Mo) and severe (Se). The soil sample were retrieved from different point, which are in the trenching area (TR), harvest path zone (H), and frond stack zone (M). The soil characteristic including microbe abundance, dehydrogenase activity, and PCR-DGGE of microbe community in each treatment were analyzed to reveal the effect of trenching construction. The results showed that trenching implementation reveals major results related to the abundance of microbes and fungi population, supported by the enhancement of dehydrogenase activity at the block with moderate Ganoderma attack. In addition, the DGGE study effectively separates the microbial population of four separate treatment into two clusters, splitting the grouping depending on the Ganoderma level attack. This study shows enhancement of soil characteristics biologically and nutrients status of oil palm leaves especially P, as the results of trenching construction in peatland. \u0000Keywords: peatland; oil palm; trenching, DGGE; microbes","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82230781","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Quality of Airborne Bacteria in Operating Theaters in Several Hospitals in Jakarta and Its Surrounding Areas in 2018-2019 2018-2019年雅加达及周边地区多家医院手术室空气细菌质量分析
Microbiology Indonesia Pub Date : 2021-03-12 DOI: 10.5454/MI.14.4.3
C. R. Tjampakasari, Nabila Naura, T. M. Sudiro
{"title":"Quality of Airborne Bacteria in Operating Theaters in Several Hospitals in Jakarta and Its Surrounding Areas in 2018-2019","authors":"C. R. Tjampakasari, Nabila Naura, T. M. Sudiro","doi":"10.5454/MI.14.4.3","DOIUrl":"https://doi.org/10.5454/MI.14.4.3","url":null,"abstract":"Nosocomial infection is an infection obtained by a patient or hospital staff while in hospital. This infectionplays a role in causing morbidity and mortality in hospitals and can occur in various hospitals rooms, includingoperating theaters. Nosocomial infections can occur due to various factors, one of which is contamination fromairborne bacteria. In some countries, regulations are set to limit the concentration of airborne bacteria, both in theoperating theaters and other rooms in hospitals, hence the need for monitoring and supervision of air quality asareflection on the cleanliness conditions in hospitals. Based on this, it is necessary to know the bacteriological airquality in the operating theaters in several hospitals in Jakarta and surrounding areas as one of the steps to preventnosocomial infections. The method uses an air sampler with the principle of impaction. Air sampler works byseparating the particles from the air by utilizing the inertia of the particles to force the bacteria to settle to thesurface of the medium. A total of 217 examinations in the operating theaters were carried out in 17 hospitals inJakarta and surrounding areas during January 2018 to June 2019. The majority of the operating theaters inhospitals in Jakarta and surrounding areas have air quality that met appropriate quality standards. In 2018, 120 of137 (87.59%) examination in the operating theaters met the quality standar. Meanwhile in 2019, 70 of 80(87.50%) operating theaters met the standard determined by the Ministry of Health of the Republic of Indonesia.\u0000Key words: air sampler method, operating theater, quality of air bacteria","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83118518","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
The Ethanol Production Activity of Indigenous Thermotolerant Yeast Pichia kudriavzevii 1P4 本土耐热酵母Pichia kudriavzevii 1P4产乙醇活性研究
Microbiology Indonesia Pub Date : 2021-02-28 DOI: 10.5454/MI.14.4.1
Darojatul Ulya, R. Astuti, A. Meryandini
{"title":"The Ethanol Production Activity of Indigenous Thermotolerant Yeast Pichia kudriavzevii 1P4","authors":"Darojatul Ulya, R. Astuti, A. Meryandini","doi":"10.5454/MI.14.4.1","DOIUrl":"https://doi.org/10.5454/MI.14.4.1","url":null,"abstract":"Pichia kudriavzevii 1P4 is a thermotolerant-ethanologenic yeast potential for application in ethanol industry. In this study we evaluated the stress tolerance phenotype of P. kudriavzevii 1P4 in dealing with fermentation related-stresses, including high temperature stress, high sugar content, ethanol content and the fermentation capacity of the particular isolate. Based on spot assay, 1P4 showed stress tolerance phenotype against high sugar concentration for up to 30% sugar content and 10% ethanol stress. In addition, 1P4 was capable to show temperature-stress tolerance phenotype for up to 42oC, suggesting that 1P4 belong to thermotolerant yeast isolate. Fermentative activity was measured by using glucose consumption and ethanol production assay. We evaluated the fermentative and growth rate of 1P4 at various temperature condition which were 27oC, 37oC and 42oC using YPD media (at initial glucose of 2%, 10% and 20%). Interestingly, 1P4 consumed the highest glucose in 20% of concentration at 37oC (15.29%), simultaneously with the highest concentration of ethanol (32.05 g/L ethanol and 0.67 g/L/h ethanol productivity). Cell growth analysis showed that growth of 1P4 isolate increased with higher initial glucose condition yet decreased as temperature during fermentation was raised. The growth rate of 1P4 was found high in 20% initial glucose at 37oC than 2% and 10% at same temperature. In addition, 1P4 exhibited short lag phase at high-temperature fermentation. Our data indicate that 1P4 can potentially be applied as fermentation agent especially in high-temperature ethanol fermentation.\u0000Key words: bioethanol production, fermentation related-stress, thermotolerant yeast","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83578385","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 4
Identification of Dermatophyte Fungi Causing Tinea pedis and Tinea unguium in Malabero Coastal Communities, Bengkulu 蚌库鲁马拉贝罗沿海社区引起足癣和龙蹄癣的皮肤真菌鉴定
Microbiology Indonesia Pub Date : 2021-01-01 DOI: 10.5454/mi.15.1.4
M. Sariyanti, Putri Maya Agustria, Willujeng Fanny Herlambang, Besly Sinuhaji, R. H. Wibowo, Novriantika Lestari, E. Nugraheni, S. Sipriyadi
{"title":"Identification of Dermatophyte Fungi Causing Tinea pedis and Tinea unguium in Malabero Coastal Communities, Bengkulu","authors":"M. Sariyanti, Putri Maya Agustria, Willujeng Fanny Herlambang, Besly Sinuhaji, R. H. Wibowo, Novriantika Lestari, E. Nugraheni, S. Sipriyadi","doi":"10.5454/mi.15.1.4","DOIUrl":"https://doi.org/10.5454/mi.15.1.4","url":null,"abstract":"","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86491120","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Exploration of Lignocellulolytic Microbes in Oil Palm Rhizosphere on Peat Soils and Their Respiration Activities 泥炭土壤油棕根际降解木质纤维素微生物及其呼吸活性研究
Microbiology Indonesia Pub Date : 2021-01-01 DOI: 10.5454/mi.15.1.5
H. Widiastuti, Bio-industry, S. Siswanto, D. Taniwiryono, Heru Bagus Pulunggono, Syaiful Anwar, B. Sumawinata, Husni Mubarok, S. Sabiham
{"title":"Exploration of Lignocellulolytic Microbes in Oil Palm Rhizosphere on Peat Soils and Their Respiration Activities","authors":"H. Widiastuti, Bio-industry, S. Siswanto, D. Taniwiryono, Heru Bagus Pulunggono, Syaiful Anwar, B. Sumawinata, Husni Mubarok, S. Sabiham","doi":"10.5454/mi.15.1.5","DOIUrl":"https://doi.org/10.5454/mi.15.1.5","url":null,"abstract":"","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80973367","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 3
Characterization of EPS7-like Enterobacteria phage Isolated from Indonesia 印度尼西亚产eps7样肠杆菌噬菌体的鉴定
Microbiology Indonesia Pub Date : 2021-01-01 DOI: 10.5454/mi.15.1.2
A. Atikana, K. Fujita, Alex Prima, Yopi Yopi, H. Kawasaki, Ken-ichiro Suzuki, P. Lisdiyanti
{"title":"Characterization of EPS7-like Enterobacteria phage Isolated from Indonesia","authors":"A. Atikana, K. Fujita, Alex Prima, Yopi Yopi, H. Kawasaki, Ken-ichiro Suzuki, P. Lisdiyanti","doi":"10.5454/mi.15.1.2","DOIUrl":"https://doi.org/10.5454/mi.15.1.2","url":null,"abstract":"Bacteriophages are the most abundant entities in earth. The order is the largest and most Caudovirales widespread group among bacterial viruses. The purpose of this study was to characterize bacteriophages from Indonesian waters. During this experiment, we collected sample from Kuningan (West Java) and Buleleng (Bali), Indonesia. We used an overlay agar method with three strains of as a host (NBRC 13965, NBRC 12713 and E. coli NBRC 13168) combined with digestion profiling using three restriction enzymes (PvuII, EcoRV and HincII) and transmission electron microscope (TEM) to characterize the morphology of the phage from Indonesia. Our results showed that phage LIPI13-Bp006 is in a group of and highly similar to Enterobacteria phage EPS7.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91424177","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Immunogenicity of Recombinant DNA Vaccine Encoding Non-Structural Protein-1 Dengue Virus Serotype-2 in Balb/c Mice 编码非结构蛋白1 -2型登革热病毒重组DNA疫苗在Balb/c小鼠中的免疫原性
Microbiology Indonesia Pub Date : 2021-01-01 DOI: 10.5454/mi.15.2.5
Fithriyah Sjatha, Elitha Sundari Pulungan, T. M. Sudiro
{"title":"Immunogenicity of Recombinant DNA Vaccine Encoding Non-Structural Protein-1 Dengue Virus Serotype-2 in Balb/c Mice","authors":"Fithriyah Sjatha, Elitha Sundari Pulungan, T. M. Sudiro","doi":"10.5454/mi.15.2.5","DOIUrl":"https://doi.org/10.5454/mi.15.2.5","url":null,"abstract":"Dengue Hemorrhagic Fever (DHF) is an infectious disease caused by the dengue virus (DENV) which spread widely in tropical and subtropical regions of the world. DENV is a single-positive strand RNA virus with a genome size of ± 11kb which encodes three structural proteins, seven non-structural proteins, and two untranslated regions (UTR). The non-structural protein-1 (NS1) of DENV is known to have important role in dengue pathogenesis also promising to be developed as dengue vaccine. Lately, novel vaccine approach by DNA immunization have given new perspective for a safe, stable, and immunogenic vaccine platform. Previously, we had successfully constructed DNA vaccine encoding NS1 protein of DENV2 (pUNS1) which expressed recombinant NS1 protein in mammalian cells line. Thus, in this current study the ability of pUNS1 to induce humoral immune response will be further analyzed by in mice immunization. Sixteen BALB/c mice aged of 4 weeks were immunized 3 times with 100 µg of pUNS1 or pUMVC4a on 2 weeks interval. Blood sampling was carried out just before immunization and termination was done 2 weeks after last immunization. Antibodies titer from individual mice sera against DENV-2 were measured with in-house ELISA. Anti-dengue NS1 IgG titer from mice group immunized with recombinant pUNS1 Showed ELISA absorbances five times higher than pUMVC4a group. This result suggested the ability of pUNS1 to induce humoral immune response against NS1 DENV-2 in-vivo. Recombinant pUNS1 can induce humoral immune response in mice.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76478808","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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