Pseudomonas fluorescens is plant growth promoting rhizobacteria (PGPR) often inoculated on plants as natural biocontrol agent capable of protecting the plants from soil-borne pathogens. Chemotactic motility allows populations of P. fluorescens to rapidly search for nutrients and is an important factor determining their competitive success to colonize plant root. Therefore, we investigated various glucose concentrations from 0% to 1% (w/v) to enhance chemotactic motility and growth of this rhizobia. Chemotactic motility was evaluated using swim plate assay and bacterial growth was measured using UV-Vis Spectrophotometer in LB and M9 medium. Glucose with low concentration (0.05%) showed to have optimum response in P. fluorescens chemotactic motility with colony diameter 38.3 mm in LB medium and 12.8 mm in M9 medium. Highest growth of P. fluorescens was 7 -1 seen in control condition of LB medium reaching a peak at 0.0246 OD (~±1.44x10 CFU mL ) while growth in 600 7 -1 M9 medium supplemented with 1% glucose was just slightly lower with 0.0227 OD (~±1.32x10 CFU mL ). 600 Glucose in high concentration showed to repress chemotactic motility and first growth phase of P. fluorescens in LB medium due to catabolite repression.
{"title":"Chemotactic Motility and Growth of Pseudomonas fluorescens Towards Glucose Concentration","authors":"Endah Rosa, U. M. Batubara, S. Suparjo","doi":"10.5454/mi.13.2.1","DOIUrl":"https://doi.org/10.5454/mi.13.2.1","url":null,"abstract":"Pseudomonas fluorescens is plant growth promoting rhizobacteria (PGPR) often inoculated on plants as natural biocontrol agent capable of protecting the plants from soil-borne pathogens. Chemotactic motility allows populations of P. fluorescens to rapidly search for nutrients and is an important factor determining their competitive success to colonize plant root. Therefore, we investigated various glucose concentrations from 0% to 1% (w/v) to enhance chemotactic motility and growth of this rhizobia. Chemotactic motility was evaluated using swim plate assay and bacterial growth was measured using UV-Vis Spectrophotometer in LB and M9 medium. Glucose with low concentration (0.05%) showed to have optimum response in P. fluorescens chemotactic motility with colony diameter 38.3 mm in LB medium and 12.8 mm in M9 medium. Highest growth of P. fluorescens was 7 -1 seen in control condition of LB medium reaching a peak at 0.0246 OD (~±1.44x10 CFU mL ) while growth in 600 7 -1 M9 medium supplemented with 1% glucose was just slightly lower with 0.0227 OD (~±1.32x10 CFU mL ). 600 Glucose in high concentration showed to repress chemotactic motility and first growth phase of P. fluorescens in LB medium due to catabolite repression.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"11 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82438080","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}
{"title":"Levels of TNF-α in PBMC (Peripheral Blood Mononuclear Cells) Induced by Recombinant Non Structural 1 Protein of Dengue Virus Serotype-2 in vitro","authors":"Fithriyah Sjatha, Oktivia CHANDRA MUSTIKA, Beti Ernawati Dewi, Tjahjani MIRAWATI SUDIRO","doi":"10.5454/mi.13.2.4","DOIUrl":"https://doi.org/10.5454/mi.13.2.4","url":null,"abstract":"","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"48 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79761306","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}
Agnes Sri Harti, N. Puspawati, Rahajeng Putriningrum
Anti-microbial bioactive compounds from snail ( Achatina fullica Ferussac) contained in snail seromucoid. It contains bioactive compounds such as glycans, peptides, glycopeptides, and chondroitin sulfate which can function as biological response modifiers (BRM) immunostimulators. Immunostimulators are compounds that can increase cellular immune responses in various ways, namely increasing the number and activity of T cells, NK cells, and macrophages and releasing interferons and interleukin. Immunostimulators are compounds that can increase cellular immune responses in various ways, namely increasing the number and activity of T cells, NK cells, macrophages and releasing interferons and interleukins.The purpose of this study was to analyze antimicrobial bioactive seromucoid compound of snail ( Achatina fullica Ferrusac) as biological response modifiers (BRM) immunostimulators. The research methods based on experimental laboratory results with research stages including snail seromucoid isolation; antimicrobial activity; characterization physicochemical and profile of snail seromucoid proteins. The results of antimicrobial activity showed that 100% seromucoid concentrations had MIC (Minimal Inhibition Concentration) in Staphylococcus aureus, Candida albicans, and Pseudomonas aeruginosa . The physicochemical examination results showed specific gravity of 1.010; pH 8, -1 -1 -1 glucose 16 mg dL ; 9 mg dL cholesterol; protein 2.8 mg dL and negative heavy metals (Pb, Cu, Hg, Al). The results of the analysis of protein profiles showed that there were 3 subunits of proteins, range from 55 to 72 kDa and 1 specific protein sub unit of 43 kDa which was thought to be antimicrobial and biological response modifiers (BRM) immunostimulators.
{"title":"Antimicrobial Bioactive Compounds of Snail Seromucoid as Biological Response Modifier Immunostimulator","authors":"Agnes Sri Harti, N. Puspawati, Rahajeng Putriningrum","doi":"10.5454/mi.13.2.3","DOIUrl":"https://doi.org/10.5454/mi.13.2.3","url":null,"abstract":"Anti-microbial bioactive compounds from snail ( Achatina fullica Ferussac) contained in snail seromucoid. It contains bioactive compounds such as glycans, peptides, glycopeptides, and chondroitin sulfate which can function as biological response modifiers (BRM) immunostimulators. Immunostimulators are compounds that can increase cellular immune responses in various ways, namely increasing the number and activity of T cells, NK cells, and macrophages and releasing interferons and interleukin. Immunostimulators are compounds that can increase cellular immune responses in various ways, namely increasing the number and activity of T cells, NK cells, macrophages and releasing interferons and interleukins.The purpose of this study was to analyze antimicrobial bioactive seromucoid compound of snail ( Achatina fullica Ferrusac) as biological response modifiers (BRM) immunostimulators. The research methods based on experimental laboratory results with research stages including snail seromucoid isolation; antimicrobial activity; characterization physicochemical and profile of snail seromucoid proteins. The results of antimicrobial activity showed that 100% seromucoid concentrations had MIC (Minimal Inhibition Concentration) in Staphylococcus aureus, Candida albicans, and Pseudomonas aeruginosa . The physicochemical examination results showed specific gravity of 1.010; pH 8, -1 -1 -1 glucose 16 mg dL ; 9 mg dL cholesterol; protein 2.8 mg dL and negative heavy metals (Pb, Cu, Hg, Al). The results of the analysis of protein profiles showed that there were 3 subunits of proteins, range from 55 to 72 kDa and 1 specific protein sub unit of 43 kDa which was thought to be antimicrobial and biological response modifiers (BRM) immunostimulators.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"18 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86122176","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}
Mohammad Rizki Fadhil Pratama, D. H. Tjahjono, T. Gusdinar
The artemisinin and its derivatives antifungal activity continue to be an interesting research object, with the potential shown to be developed as an antifungal compound. Artesunate, one of the artemisinin derivatives known to have antifungal activity against various pathogenic fungi, including Candida albicans. This study aims to determine the effect of artesunate on antifungal activity toward C. albicans in vitro at concentrations below 1 mgmL-1. The method used is yeast-plate count, with a parameter of observation were the number of C. albicans colonies viable after exposure with artesunate for five days. The concentration of artesunate used was divided into six groups, which were 10; 1; 10-1; 10-2; 10-3; and 10-4mgmL-1. Compared to control, a significant decrease in colony counts was only shown at the highest concentration of 10 mgmL-1. Interestingly, at the lowest concentration of 10-4mgmL-1, it showed an increase in a number of colonies almost twice of the blank. These results suggest that while at higher concentration of artesunate may inhibit the growth of C. albicans, a lower concentration of artesunate may stimulate their growth.
{"title":"The Antifungal Activity of Artesunate toward Candida albicans: Two Opposite Activities","authors":"Mohammad Rizki Fadhil Pratama, D. H. Tjahjono, T. Gusdinar","doi":"10.5454/MI.13.3.5","DOIUrl":"https://doi.org/10.5454/MI.13.3.5","url":null,"abstract":"The artemisinin and its derivatives antifungal activity continue to be an interesting research object, with the potential shown to be developed as an antifungal compound. Artesunate, one of the artemisinin derivatives known to have antifungal activity against various pathogenic fungi, including Candida albicans. This study aims to determine the effect of artesunate on antifungal activity toward C. albicans in vitro at concentrations below 1 mgmL-1. The method used is yeast-plate count, with a parameter of observation were the number of C. albicans colonies viable after exposure with artesunate for five days. The concentration of artesunate used was divided into six groups, which were 10; 1; 10-1; 10-2; 10-3; and 10-4mgmL-1. Compared to control, a significant decrease in colony counts was only shown at the highest concentration of 10 mgmL-1. Interestingly, at the lowest concentration of 10-4mgmL-1, it showed an increase in a number of colonies almost twice of the blank. These results suggest that while at higher concentration of artesunate may inhibit the growth of C. albicans, a lower concentration of artesunate may stimulate their growth.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89320104","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}
Radish ( Raphanus sativus L.) root is commonly used as flavor enhancing additive or side dish. Previous research revealed the presence of active compound in which could inhibit bacterial growth. Thus, a research concerning natural antibacterial for fish products that are categorized as high-risk food being contaminated by spoilage bacteria ( Pseudomonas aeruginosa, Bacillus cereus, dan Staphylococcus aureus ) was done. Radish root extraction was made by using ethyl acetate (semi polar) for 3 days. Well diffusion was performed using 4 extract concentration (10, 20, 30, dan 40% (w/v)) againts three fish spoilage bacteria. Based on our results, 30% concentration was the best concentration which inhibit more than 10 mm in inhibition zone with MIC and MBC. The scores were of 0.06% and 0.24% ( P. aeruginosa ), 0.13% and 0.50% ( S. aureus ), and 0.12% and 0.48% ( B. cereus ). Moreover, based on stability test against heating temperature showed that this extract concentration was more stable in 80°C with duration times for 5 minutes and pH 3 which resulting the lowest inhibition zone reduction compares to control extract. Radish extract was categorized as low toxic compound (LC 50 = 839.52 ppm) functioning in antibacterial compound containing major antibacterial compound ( bis(2-ethylhexyl) phthalate , 1,2- benzenedicarboxylic acid , 9,12,15-octadecatrienoic acid ), fatty acid ( n-hexadecanoic acid , butanedioic acid ), carboxylic acid ( isobutyric acid , malic acid , oleic acid ), and minor antibacterial compound ( n-Hydroxymethylacetamide , 2,4-bis(1,1-dimethylethyl) , 2,4-pentanedione , 2-Cyclohexen-1-one , hydrazine , cyclohexene oxide , gamma-sitosterol ).
{"title":"Antibacterial Potential of Radish Extract (Raphanus sativus L.) Against Fish Spoilage Bacteria","authors":"Eveline Eveline, Chikita Wini Tanumihardja","doi":"10.5454/MI.13.3.3","DOIUrl":"https://doi.org/10.5454/MI.13.3.3","url":null,"abstract":"Radish ( Raphanus sativus L.) root is commonly used as flavor enhancing additive or side dish. Previous research revealed the presence of active compound in which could inhibit bacterial growth. Thus, a research concerning natural antibacterial for fish products that are categorized as high-risk food being contaminated by spoilage bacteria ( Pseudomonas aeruginosa, Bacillus cereus, dan Staphylococcus aureus ) was done. Radish root extraction was made by using ethyl acetate (semi polar) for 3 days. Well diffusion was performed using 4 extract concentration (10, 20, 30, dan 40% (w/v)) againts three fish spoilage bacteria. Based on our results, 30% concentration was the best concentration which inhibit more than 10 mm in inhibition zone with MIC and MBC. The scores were of 0.06% and 0.24% ( P. aeruginosa ), 0.13% and 0.50% ( S. aureus ), and 0.12% and 0.48% ( B. cereus ). Moreover, based on stability test against heating temperature showed that this extract concentration was more stable in 80°C with duration times for 5 minutes and pH 3 which resulting the lowest inhibition zone reduction compares to control extract. Radish extract was categorized as low toxic compound (LC 50 = 839.52 ppm) functioning in antibacterial compound containing major antibacterial compound ( bis(2-ethylhexyl) phthalate , 1,2- benzenedicarboxylic acid , 9,12,15-octadecatrienoic acid ), fatty acid ( n-hexadecanoic acid , butanedioic acid ), carboxylic acid ( isobutyric acid , malic acid , oleic acid ), and minor antibacterial compound ( n-Hydroxymethylacetamide , 2,4-bis(1,1-dimethylethyl) , 2,4-pentanedione , 2-Cyclohexen-1-one , hydrazine , cyclohexene oxide , gamma-sitosterol ).","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"35 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89737712","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}
L. Lindayani, Laksmi Hartajanie, Monika Palupi Murniati
Yellow bamboo shoot is a popular food material especially in Semarang because it is used as filling for lumpia (authentic food of Semarang). Beside used as filling, yellow bamboo shoot commonly known by Indonesian people. Considering bamboo shoot often processed into fermented traditional food, yellow bamboo shoot also potential to be examined as source of fermented food producing lactic acid bacteria. Lactic acid bacteria still become the most beneficial microorganisms associated with gastrointestinal system and moreover for obesity diet. The aim of this research is to study the probiotic potential of lactic acid bacteria produced from yellow bamboo shoot fermentation in 2.5% and 5% brine under room temperature (30 °C). From isolation, 22 single colonies obtained from 2.5% brine and 27 isolates obtained from 5% brine. The morphology and physiology analysis resulted in Lactobacillus and Streptococcus genus. All isolates were tested subsequently for probiotic potential. Based on the result, more than 50% identified isolates have probiotic potential.
{"title":"Probiotic Potential of Lactic Acid Bacteria from Yellow Bamboo Shoot Fermentation using 2.5% and 5% Brine at Room Temperature","authors":"L. Lindayani, Laksmi Hartajanie, Monika Palupi Murniati","doi":"10.5454/mi.12.1.5","DOIUrl":"https://doi.org/10.5454/mi.12.1.5","url":null,"abstract":"Yellow bamboo shoot is a popular food material especially in Semarang because it is used as filling for lumpia (authentic food of Semarang). Beside used as filling, yellow bamboo shoot commonly known by Indonesian people. Considering bamboo shoot often processed into fermented traditional food, yellow bamboo shoot also potential to be examined as source of fermented food producing lactic acid bacteria. Lactic acid bacteria still become the most beneficial microorganisms associated with gastrointestinal system and moreover for obesity diet. The aim of this research is to study the probiotic potential of lactic acid bacteria produced from yellow bamboo shoot fermentation in 2.5% and 5% brine under room temperature (30 °C). From isolation, 22 single colonies obtained from 2.5% brine and 27 isolates obtained from 5% brine. The morphology and physiology analysis resulted in Lactobacillus and Streptococcus genus. All isolates were tested subsequently for probiotic potential. Based on the result, more than 50% identified isolates have probiotic potential.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"51 1","pages":"5"},"PeriodicalIF":0.0,"publicationDate":"2018-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77852773","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}
V. A. Testamenti, D. Iskandriati, A. Wahyudi, J. Pamungkas
Melioidosis is an emerging zoonotic disease caused by the saprophytic Burkholderia pseudomallei, which infects human and various range of animal species. Septicaemia and pneumonia in human patients often lead to death when the patient is not treated. The disease is spread in tropical areas, especially in Southeast Asia and Northern Australia, but in Indonesia the cases are poorly reported. This research provides serological evidence of antibodies to B. pseudomallei in both captive and wild nonhuman primates captured by enzyme-linked immunosorbent assay (ELISA). Plasma samples were taken from a total of 390 monkeys both in captivities and in wild habitat. The seroprevalence varies from 11%-80% . The findings showed that infection by B. pseudomallei in both captive and wild macaques has reached an alarming level and requires further studies.
{"title":"Detection of Antibody to Burkholderia pseudomallei in Captive and Wild Macaques","authors":"V. A. Testamenti, D. Iskandriati, A. Wahyudi, J. Pamungkas","doi":"10.5454/MI.1.1.%P","DOIUrl":"https://doi.org/10.5454/MI.1.1.%P","url":null,"abstract":"Melioidosis is an emerging zoonotic disease caused by the saprophytic Burkholderia pseudomallei, which infects human and various range of animal species. Septicaemia and pneumonia in human patients often lead to death when the patient is not treated. The disease is spread in tropical areas, especially in Southeast Asia and Northern Australia, but in Indonesia the cases are poorly reported. This research provides serological evidence of antibodies to B. pseudomallei in both captive and wild nonhuman primates captured by enzyme-linked immunosorbent assay (ELISA). Plasma samples were taken from a total of 390 monkeys both in captivities and in wild habitat. The seroprevalence varies from 11%-80% . The findings showed that infection by B. pseudomallei in both captive and wild macaques has reached an alarming level and requires further studies.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74707373","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}
V. A. Testamenti, D. Iskandriati, A. Wahyudi, J. Pamungkas
Melioidosis is an emerging zoonotic disease caused by the saprophytic Burkholderia pseudomallei, which infects human and a wide range of animal species. Melioidosis may lead to septicemia and pneumonia in human patients, which can be fatal if the patient is not treated accordingly. The disease is spread in tropical areas and is highly endemic to Southeast Asia and Northern Australia. However, melioidosis is poorly reported in Indonesia, especially in the veterinary field. This research provides serological evidence of antibodies to B. pseudomallei in both captive and wild nonhuman primates. Plasma samples were taken from a total of 390 monkeys in captivities and wild habitats in West Java and Bali, Indonesia. Enzyme-linked immunosorbent assay (ELISA) showed that the seroprevalence was 42.21% for Macaca fascicularis and 43.59% for Macaca nemestrina. Furthermore, the seroprevalence was 53.41% for captive macaques and 17.83% for wild macaques. The findings showed that exposure to B. pseudomallei happened in both captive and wild macaques. Based on this serosurveillance results, further studies such as comprehensive culture and clinical study are required to discover the clinical burden of the disease in nonhuman primates.
{"title":"Detection of Antibody to Burkholderia pseudomallei in Captive and Wild Macaquesin West Java and Bali, Indonesia","authors":"V. A. Testamenti, D. Iskandriati, A. Wahyudi, J. Pamungkas","doi":"10.5454/MI.12.1.4","DOIUrl":"https://doi.org/10.5454/MI.12.1.4","url":null,"abstract":"Melioidosis is an emerging zoonotic disease caused by the saprophytic Burkholderia pseudomallei, which infects human and a wide range of animal species. Melioidosis may lead to septicemia and pneumonia in human patients, which can be fatal if the patient is not treated accordingly. The disease is spread in tropical areas and is highly endemic to Southeast Asia and Northern Australia. However, melioidosis is poorly reported in Indonesia, especially in the veterinary field. This research provides serological evidence of antibodies to B. pseudomallei in both captive and wild nonhuman primates. Plasma samples were taken from a total of 390 monkeys in captivities and wild habitats in West Java and Bali, Indonesia. Enzyme-linked immunosorbent assay (ELISA) showed that the seroprevalence was 42.21% for Macaca fascicularis and 43.59% for Macaca nemestrina. Furthermore, the seroprevalence was 53.41% for captive macaques and 17.83% for wild macaques. The findings showed that exposure to B. pseudomallei happened in both captive and wild macaques. Based on this serosurveillance results, further studies such as comprehensive culture and clinical study are required to discover the clinical burden of the disease in nonhuman primates.","PeriodicalId":18546,"journal":{"name":"Microbiology Indonesia","volume":"8 1","pages":"4-4"},"PeriodicalIF":0.0,"publicationDate":"2018-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75435929","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}
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