Pub Date : 2024-07-19eCollection Date: 2024-01-01DOI: 10.1155/2024/6251407
Joyce Maria Schuch, Carolina Rosai Mendes, Guilherme Lopes Cardoso, Carlos André da Veiga Lima Rosa Costamilan, Paulo Renato Matos Lopes, Renato Nallin Montagnolli, Guilherme Dilarri, Ederio Dino Bidoia
The fungus Phyllosticta citricarpa is a quarantine phytopathogen responsible for causing citrus black spot (CBS) disease. To export fruits to CBS-free countries, they must undergo a sanitation process to ensure disease control. In this study, neem essential oil (NEO) was tested against P. citricarpa for the first time as an alternative sanitizer. In vitro experiments were conducted to determine the inhibition concentration of NEO for P. citricarpa, and the mode of action of the essential oil was evaluated. In vivo assays were performed to simulate the sanitization process used in packinghouses. NEO was characterized by GC-MS/MS. The results revealed that NEO at 100 μL·mL-1 exhibited a similar inhibitory effect as copper oxychloride, suppressing 89.68 ± 1.14% of fungal mycelium growth. Fluorescence microscopy experiments demonstrated that NEO functions by disrupting the cytoplasmic membrane of fungal hyphae, leading to their death within 30 minutes of contact with NEO. GC-MS/MS characterization revealed a high presence of phenolic compounds, which serve as the primary antifungal agents responsible for the action against fungal hyphae. In vivo assays showed that NEO at 100 μL·mL-1 also reduced microorganisms (CFU mL-1) by 93.00 ± 3.88% compared to the negative control. Overall, the results demonstrate that NEO can effectively serve as an alternative sanitizer against P. citricarpa in citrus packinghouses. Our findings allow future studies to explore the use of NEO for sanitizing other fruits and combating different phytopathogens to broaden its potential application in fruit sanitation for export.
{"title":"Neem Essential Oil as an Antifungal Agent against <i>Phyllosticta citricarpa</i>.","authors":"Joyce Maria Schuch, Carolina Rosai Mendes, Guilherme Lopes Cardoso, Carlos André da Veiga Lima Rosa Costamilan, Paulo Renato Matos Lopes, Renato Nallin Montagnolli, Guilherme Dilarri, Ederio Dino Bidoia","doi":"10.1155/2024/6251407","DOIUrl":"10.1155/2024/6251407","url":null,"abstract":"<p><p>The fungus <i>Phyllosticta citricarpa</i> is a quarantine phytopathogen responsible for causing citrus black spot (CBS) disease. To export fruits to CBS-free countries, they must undergo a sanitation process to ensure disease control. In this study, neem essential oil (NEO) was tested against <i>P. citricarpa</i> for the first time as an alternative sanitizer. <i>In vitro</i> experiments were conducted to determine the inhibition concentration of NEO for <i>P. citricarpa</i>, and the mode of action of the essential oil was evaluated. <i>In vivo</i> assays were performed to simulate the sanitization process used in packinghouses. NEO was characterized by GC-MS/MS. The results revealed that NEO at 100 <i>μ</i>L·mL<sup>-1</sup> exhibited a similar inhibitory effect as copper oxychloride, suppressing 89.68 ± 1.14% of fungal mycelium growth. Fluorescence microscopy experiments demonstrated that NEO functions by disrupting the cytoplasmic membrane of fungal hyphae, leading to their death within 30 minutes of contact with NEO. GC-MS/MS characterization revealed a high presence of phenolic compounds, which serve as the primary antifungal agents responsible for the action against fungal hyphae. <i>In vivo</i> assays showed that NEO at 100 <i>μ</i>L·mL<sup>-1</sup> also reduced microorganisms (CFU mL<sup>-1</sup>) by 93.00 ± 3.88% compared to the negative control. Overall, the results demonstrate that NEO can effectively serve as an alternative sanitizer against <i>P. citricarpa</i> in citrus packinghouses. Our findings allow future studies to explore the use of NEO for sanitizing other fruits and combating different phytopathogens to broaden its potential application in fruit sanitation for export.</p>","PeriodicalId":14098,"journal":{"name":"International Journal of Microbiology","volume":"2024 ","pages":"6251407"},"PeriodicalIF":2.8,"publicationDate":"2024-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11281856/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141788005","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-19eCollection Date: 2024-01-01DOI: 10.1155/2024/8177437
Mohammed Jasim Mohammed, Abbas S Al-Mizraqchi, Salah M Ibrahim
Background: Type 2 diabetes is a condition in which the body becomes resistant to the effects of insulin, leading to reduced insulin production in the pancreas. It has genetic- and family-related risk factors that cannot be changed, along with modifiable lifestyle factors. The precise genetic causes of type 2 diabetes are still unknown. However, individuals can potentially slow or stop the progression of the condition by making dietary adjustments and increasing physical activity levels. Material and Methods. Forty-five type II diabetic patients in the study included participants between 40 and 60 years old, with a minimum duration of one year, as well as 45 healthy control subjects who were matched in terms of age and sex, and had no underlying systemic diseases. Oral examination is done for the symptoms including burning sensation, candidiasis, and a reduction in the production of saliva. The rate of saliva flow (in milliliters per minute) was measured in samples of saliva that were not stimulated. The salivary trace elements and levels of adipocytokines were evaluated using colorimetric and Ethylenediaminetetraacetic acid (ELISA) testing. The quantification of Candida colony numbers, an enrichment and culture approach, was used to achieve a concentration of 100,000 colony-forming units per milliliter (CFU/ml). The ShowNovo WG1 halimeter was used to measure volatile sulfur compounds in breath. The salivary glucose oxidase assay was conducted using a colorimetric technique, while the determination of trace elements was also performed using a colorimetric assay method.
Result: The diabetic group exhibited a significant increase in the number of Candida spp colonies due to elevated levels of glucose in the saliva (p > 0.05). However, the variables being examined, such as body mass index (BMI), burning mouth syndrome (BMS), salivary flow rate (SFR), salivary leptin, salivary copper, and salivary magnesium, did not exhibit any significant variations in quantities between the diabetic and healthy groups (p > 0.05).
Conclusion: The data collected in this research aid in the creation of a preventative program for oral fungal infections in individuals with type 2 diabetes. The program utilizes saliva and its constituents.
{"title":"Oral Findings, Salivary Copper, Magnesium, and Leptin in Type II Diabetic Patients in Relation to Oral <i>Candida</i> Species.","authors":"Mohammed Jasim Mohammed, Abbas S Al-Mizraqchi, Salah M Ibrahim","doi":"10.1155/2024/8177437","DOIUrl":"10.1155/2024/8177437","url":null,"abstract":"<p><strong>Background: </strong>Type 2 diabetes is a condition in which the body becomes resistant to the effects of insulin, leading to reduced insulin production in the pancreas. It has genetic- and family-related risk factors that cannot be changed, along with modifiable lifestyle factors. The precise genetic causes of type 2 diabetes are still unknown. However, individuals can potentially slow or stop the progression of the condition by making dietary adjustments and increasing physical activity levels. <i>Material and Methods</i>. Forty-five type II diabetic patients in the study included participants between 40 and 60 years old, with a minimum duration of one year, as well as 45 healthy control subjects who were matched in terms of age and sex, and had no underlying systemic diseases. Oral examination is done for the symptoms including burning sensation, candidiasis, and a reduction in the production of saliva. The rate of saliva flow (in milliliters per minute) was measured in samples of saliva that were not stimulated. The salivary trace elements and levels of adipocytokines were evaluated using colorimetric and Ethylenediaminetetraacetic acid (ELISA) testing. The quantification of <i>Candida</i> colony numbers, an enrichment and culture approach, was used to achieve a concentration of 100,000 colony-forming units per milliliter (CFU/ml). The ShowNovo WG1 halimeter was used to measure volatile sulfur compounds in breath. The salivary glucose oxidase assay was conducted using a colorimetric technique, while the determination of trace elements was also performed using a colorimetric assay method.</p><p><strong>Result: </strong>The diabetic group exhibited a significant increase in the number of <i>Candida</i> spp colonies due to elevated levels of glucose in the saliva (<i>p</i> > 0.05). However, the variables being examined, such as body mass index (BMI), burning mouth syndrome (BMS), salivary flow rate (SFR), salivary leptin, salivary copper, and salivary magnesium, did not exhibit any significant variations in quantities between the diabetic and healthy groups (<i>p</i> > 0.05).</p><p><strong>Conclusion: </strong>The data collected in this research aid in the creation of a preventative program for oral fungal infections in individuals with type 2 diabetes. The program utilizes saliva and its constituents.</p>","PeriodicalId":14098,"journal":{"name":"International Journal of Microbiology","volume":"2024 ","pages":"8177437"},"PeriodicalIF":2.8,"publicationDate":"2024-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11281854/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141788006","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Background: Bacterial bloodstream infections (BSIs) are the leading cause of sepsis-related morbidity and mortality worldwide. The emergence and spread of antimicrobial resistance (AMR) in bacteria is also a growing global concern. As a result, data on bacterial profile and their antibiogram are essential for strategies to contain drug resistance, improve the quality of patient care, and strengthen health systems.
Methods: Retrospective data from bacteriological results of blood samples of BSI-suspected patients from 2018 to 2021 were collected using a data collection sheet. Standard bacteriological techniques were followed during sample collection, culture preparation, bacterial identification, and antibiotic susceptibility testing (AST). We used Epi Info version 7 to enter and clean the data and then exported it to SPSS version 26 for analysis. Logistic regression models were used to measure the association between variables. A p value <0.05 with a 95% confidence interval was considered as statistically significant.
Result: Of the total 2,795 blood culture records, 455 (16.3%) were culture positive for bacteria, with Klebsiella pneumoniae (26%) and Staphylococcus aureus (24.6%) being the leading isolates. The isolates were highly resistant to common antibiotics, with more than 80% of them being resistant to ceftriaxone and penicillin. Moreover, about 43% of isolates were multidrug resistant (MDR), with Klebsiella pneumoniae (65.5%), Acinetobacter species (56.7%), and Citrobacter species (53.8%) being the most common MDR isolates. Age and diagnosis year were significantly associated with the presence of bacterial BSIs (p value <0.05).
Conclusion: Bacterial BSI and AMR were growing concerns in the study area. Bacteremia was more common in children under the age of five, and it decreased as the patient's age increased. The alarming rate of AMR, such as MDR blood isolates, calls for periodic and continuous monitoring of antibiotic usage in the study area.
{"title":"Antibiogram of Bacteria Isolated from Bloodstream Infection-Suspected Patients at the University of Gondar Comprehensive Specialized Hospital in Northwest Ethiopia: A Retrospective Study.","authors":"Minichil Worku, Tigist Molla, Desie Kasew, Muluneh Assefa, Alene Geteneh, Melak Aynalem, Mucheye Gizachew, Sirak Biset","doi":"10.1155/2024/7624416","DOIUrl":"10.1155/2024/7624416","url":null,"abstract":"<p><strong>Background: </strong>Bacterial bloodstream infections (BSIs) are the leading cause of sepsis-related morbidity and mortality worldwide. The emergence and spread of antimicrobial resistance (AMR) in bacteria is also a growing global concern. As a result, data on bacterial profile and their antibiogram are essential for strategies to contain drug resistance, improve the quality of patient care, and strengthen health systems.</p><p><strong>Methods: </strong>Retrospective data from bacteriological results of blood samples of BSI-suspected patients from 2018 to 2021 were collected using a data collection sheet. Standard bacteriological techniques were followed during sample collection, culture preparation, bacterial identification, and antibiotic susceptibility testing (AST). We used Epi Info version 7 to enter and clean the data and then exported it to SPSS version 26 for analysis. Logistic regression models were used to measure the association between variables. A <i>p</i> value <0.05 with a 95% confidence interval was considered as statistically significant.</p><p><strong>Result: </strong>Of the total 2,795 blood culture records, 455 (16.3%) were culture positive for bacteria, with <i>Klebsiella pneumoniae</i> (26%) and <i>Staphylococcus aureus</i> (24.6%) being the leading isolates. The isolates were highly resistant to common antibiotics, with more than 80% of them being resistant to ceftriaxone and penicillin. Moreover, about 43% of isolates were multidrug resistant (MDR), with <i>Klebsiella pneumoniae</i> (65.5%), <i>Acinetobacter</i> species (56.7%), and <i>Citrobacter</i> species (53.8%) being the most common MDR isolates. Age and diagnosis year were significantly associated with the presence of bacterial BSIs (<i>p</i> value <0.05).</p><p><strong>Conclusion: </strong>Bacterial BSI and AMR were growing concerns in the study area. Bacteremia was more common in children under the age of five, and it decreased as the patient's age increased. The alarming rate of AMR, such as MDR blood isolates, calls for periodic and continuous monitoring of antibiotic usage in the study area.</p>","PeriodicalId":14098,"journal":{"name":"International Journal of Microbiology","volume":"2024 ","pages":"7624416"},"PeriodicalIF":2.8,"publicationDate":"2024-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11250713/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141626657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-30eCollection Date: 2024-01-01DOI: 10.1155/2024/2748842
Nallely S Badillo-Larios, Edgar Alejandro Turrubiartes-Martínez, Esther Layseca-Espinosa, Roberto González-Amaro, Luis Fernando Pérez-González, Perla Niño-Moreno
Pseudomonas aeruginosa is an opportunistic pathogen in HAIs with two facets: the most studied is the high rate of antimicrobial resistance, and the less explored is the long list of virulence factors it possesses. This study aimed to characterize the virulence genes carried by strains as well as the profile of cytokines related to inflammation, according to the resistance profile presented. This study aims to identify the virulence factors associated with MDR strains, particularly those resistant to carbapenems, and assess whether there is a cytokine profile that correlates with these characteristics. As methodology species were identified by classical microbiological techniques and confirmed by molecular biology, resistance levels were determined by the minimum inhibitory concentration and identification of MDR strains. Virulence factor genotyping was performed using PCR. In addition, biofilm production was assessed using crystal violet staining. Finally, the strains were cocultured with PBMC, and cell survival and the cytokines IL-1β, IL-6, IL-10, IL-8, and TNF-α were quantified using flow cytometry. Bacteremia and nosocomial pneumonia in adults are the most frequent types of infection. In the toxigenic aspect, genes corresponding to the type III secretion system were present in at least 50% of cases. In addition, PBMC exposed to strains of four different categories according to their resistance and toxicity showed a differential pattern of cytokine expression, a decrease in IL-10, IL-6, and IL-8, and an over-secretion of IL-1b. In conclusion, the virulence genes showed a differentiated appearance for the two most aggressive exotoxins of T3SS (exoU and exoS) in multidrug-resistant strains. Moreover, the cytokine profile displays a low expression of cytokines with anti-inflammatory and proinflammatory effects in strains carrying the exoU gene.
{"title":"Interesting Cytokine Profile Caused by Clinical Strains of <i>Pseudomonas aeruginosa</i> MDR Carrying the exoU Gene.","authors":"Nallely S Badillo-Larios, Edgar Alejandro Turrubiartes-Martínez, Esther Layseca-Espinosa, Roberto González-Amaro, Luis Fernando Pérez-González, Perla Niño-Moreno","doi":"10.1155/2024/2748842","DOIUrl":"10.1155/2024/2748842","url":null,"abstract":"<p><p><i>Pseudomonas aeruginosa</i> is an opportunistic pathogen in HAIs with two facets: the most studied is the high rate of antimicrobial resistance, and the less explored is the long list of virulence factors it possesses. This study aimed to characterize the virulence genes carried by strains as well as the profile of cytokines related to inflammation, according to the resistance profile presented. This study aims to identify the virulence factors associated with MDR strains, particularly those resistant to carbapenems, and assess whether there is a cytokine profile that correlates with these characteristics. As methodology species were identified by classical microbiological techniques and confirmed by molecular biology, resistance levels were determined by the minimum inhibitory concentration and identification of MDR strains. Virulence factor genotyping was performed using PCR. In addition, biofilm production was assessed using crystal violet staining. Finally, the strains were cocultured with PBMC, and cell survival and the cytokines IL-1<i>β</i>, IL-6, IL-10, IL-8, and TNF-<i>α</i> were quantified using flow cytometry. Bacteremia and nosocomial pneumonia in adults are the most frequent types of infection. In the toxigenic aspect, genes corresponding to the type III secretion system were present in at least 50% of cases. In addition, PBMC exposed to strains of four different categories according to their resistance and toxicity showed a differential pattern of cytokine expression, a decrease in IL-10, IL-6, and IL-8, and an over-secretion of IL-1b. In conclusion, the virulence genes showed a differentiated appearance for the two most aggressive exotoxins of T3SS (<i>exoU</i> and <i>exoS</i>) in multidrug-resistant strains. Moreover, the cytokine profile displays a low expression of cytokines with anti-inflammatory and proinflammatory effects in strains carrying the exoU gene.</p>","PeriodicalId":14098,"journal":{"name":"International Journal of Microbiology","volume":"2024 ","pages":"2748842"},"PeriodicalIF":2.8,"publicationDate":"2024-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11227949/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141554742","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The common bean (Phaseolus vulgaris L.) is a yearly herbaceous plant grown for its edible dry seeds. Despite that, pests and diseases have contributed to the decline of common bean production in Kenya. Therefore, the study aimed to identify bacteria from Lake Bogoria, assess the pathogenicity of Rhizoctonia solani Kühn, screen for effective antifungal agents, and determine secondary metabolites for the biocontrol of R. solani. A total of 49 bacteria were isolated, of which 10 isolates had varied mycelial inhibition rates of R. solani in the co-culture technique. The efficacy of volatile compounds of the three selected bacterial strains had varied mycelial growth and percent reduction against R. solani. The pathogenicity assay showed varied plant parameters and biomass of R. solani on common bean plantlets. The molecular characterization based on 16 S ribosomal RNA confirmed the selected bacterial strains' identity with a diversity similar to the Bacillus genus. Gas chromatography-mass spectrometry analysis of secondary metabolites showed different antimicrobial compounds produced by Bacillus subtilis strain TW21. In conclusion, Lake Bogoria harbors useful microbes as biocontrol agents against plant pathogens. The current study discovers the potential biocontrol bacteria isolates from Lake Bogoria as alternative bioagents against R. solani. Therefore, the isolate Bacillus subtilis strain TW21 can be assessed further for toxicological and ecotoxicological studies and registered by the Pest Control Products Board (PCPB), Kenya, as a biocontrol product against common diseases affecting common beans' production.
蚕豆(Phaseolus vulgaris L.)是一种一年生草本植物,因其可食用的干种子而被种植。尽管如此,病虫害还是导致肯尼亚蚕豆产量下降。因此,本研究旨在鉴定博戈里亚湖中的细菌,评估 Rhizoctonia solani Kühn 的致病性,筛选有效的抗真菌剂,并确定用于 R. solani 生物防治的次生代谢物。共分离出 49 种细菌,其中 10 种细菌在共培养技术中对 R. solani 的菌丝抑制率各不相同。所选的三种细菌菌株的挥发性化合物对 R. solani 的菌丝生长和抑制率的功效各不相同。致病性测定显示,R. solani 在蚕豆小苗上的植物参数和生物量各不相同。基于 16 S 核糖体 RNA 的分子鉴定证实了所选细菌菌株的身份,其多样性与芽孢杆菌属相似。次生代谢物的气相色谱-质谱分析表明,枯草芽孢杆菌 TW21 菌株产生了不同的抗菌化合物。总之,博戈里亚湖蕴藏着可作为生物控制剂对抗植物病原体的有益微生物。目前的研究发现了从博格利亚湖中分离出的潜在生物防治细菌,可作为对抗 R. solani 的替代生物制剂。因此,可以对分离的枯草芽孢杆菌 TW21 菌株进行进一步的毒理学和生态毒理学研究评估,并在肯尼亚害虫控制产品委员会(PCPB)注册,作为生物防治产品,防治影响普通豆类生产的常见病。
{"title":"Study of Pathogenicity Test, Antifungal Activity, and Secondary Metabolites of <i>Bacillus</i> spp. from Lake Bogoria as Biocontrol of <i>Rhizoctonia solani</i> Kühn in <i>Phaseolu</i>s <i>vulgaris</i> L.","authors":"Tofick Barasa Wekesa, Vitalis Wafula Wekesa, Justus Mong'are Onguso, Ndinda Kavesu, Patrick Wafula Okanya","doi":"10.1155/2024/6620490","DOIUrl":"10.1155/2024/6620490","url":null,"abstract":"<p><p>The common bean (<i>Phaseolus vulgaris</i> L.) is a yearly herbaceous plant grown for its edible dry seeds. Despite that, pests and diseases have contributed to the decline of common bean production in Kenya. Therefore, the study aimed to identify bacteria from Lake Bogoria, assess the pathogenicity of <i>Rhizoctonia solani</i> Kühn, screen for effective antifungal agents, and determine secondary metabolites for the biocontrol of <i>R. solani.</i> A total of 49 bacteria were isolated, of which 10 isolates had varied mycelial inhibition rates of <i>R. solani</i> in the co-culture technique. The efficacy of volatile compounds of the three selected bacterial strains had varied mycelial growth and percent reduction against <i>R. solani</i>. The pathogenicity assay showed varied plant parameters and biomass of <i>R. solani</i> on common bean plantlets. The molecular characterization based on 16 S ribosomal RNA confirmed the selected bacterial strains' identity with a diversity similar to the <i>Bacillus</i> genus. Gas chromatography-mass spectrometry analysis of secondary metabolites showed different antimicrobial compounds produced by <i>Bacillus subtilis</i> strain TW21. In conclusion, Lake Bogoria harbors useful microbes as biocontrol agents against plant pathogens. The current study discovers the potential biocontrol bacteria isolates from Lake Bogoria as alternative bioagents against <i>R. solani</i>. Therefore, the isolate <i>Bacillus subtilis</i> strain TW21 can be assessed further for toxicological and ecotoxicological studies and registered by the Pest Control Products Board (PCPB), Kenya, as a biocontrol product against common diseases affecting common beans' production.</p>","PeriodicalId":14098,"journal":{"name":"International Journal of Microbiology","volume":"2024 ","pages":"6620490"},"PeriodicalIF":2.8,"publicationDate":"2024-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11226341/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141554743","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-11eCollection Date: 2024-01-01DOI: 10.1155/2024/7685878
Michael F Kengne, Armelle T Mbaveng, Ousenu Karimo, Ballue S T Dadjo, Ornella D Tsobeng, Wiliane J T Marbou, Victor Kuete
Background. Opportunistic infections are the second cause of death among cancer patients. This study aimed at determining the antimicrobial profile and the prevalence of extended-spectrum beta-lactamase (ESBL)-gene carriage of Pseudomonas aeruginosa isolates among cancer patients at the Douala Laquintinie Hospital, Littoral Region of Cameroon. Between October 2021 and March 2023, 507 study participants were recruited among whom 307 (60.55%) were cancer patients, compared to 200 (39.45%) noncancer patients. Fifty-eight P. aeruginosa isolates were isolated from fecal samples of forty-five cancer patients and thirteen noncancer patients using Cetrimide agar. The antimicrobial resistance profile of the isolates was determined using the Kirby-Bauer disk diffusion method. The polymerase chain reaction was used to detect the presence of extended-spectrum beta-lactamase genes among P. aeruginosa isolates. P. aeruginosa showed significant resistance rates in cancer patients compared to noncancer patients to imipenem, cefotaxime, and ceftazidime, piperacillin-tazobactam, ticarcillin-clavulanic acid, and ciprofloxacin. The multidrug resistance (MDR) rate was significantly (p < 0.05) higher in cancer patients than in noncancer patients. The frequency of beta-lactamase genes in the 58 ESBL-producing P. aeruginosa isolates was determined as 72.41% for blaTEM, 37.93% for blaOXA, 74.14% for blaCTX-M, and 44.83% for blaSHV genes. The study revealed an alarmingly high prevalence of fecal carriage of ESBL-producing P. aeruginosa with a high rate of MDR among cancer patients. It indicates that regular monitoring and surveillance of ESBL-producing P. aeruginosa among cancer patients are needed to improve the management of patients.
{"title":"Frequency of Fecal Carriage of ESBL Resistance Genes in Multidrug-Resistant <i>Pseudomonas aeruginosa</i> Isolates from Cancer Patients at Laquintinie Hospital, Douala, Littoral Region, Cameroon.","authors":"Michael F Kengne, Armelle T Mbaveng, Ousenu Karimo, Ballue S T Dadjo, Ornella D Tsobeng, Wiliane J T Marbou, Victor Kuete","doi":"10.1155/2024/7685878","DOIUrl":"10.1155/2024/7685878","url":null,"abstract":"<p><p><i>Background</i>. Opportunistic infections are the second cause of death among cancer patients. This study aimed at determining the antimicrobial profile and the prevalence of extended-spectrum beta-lactamase (ESBL)-gene carriage of <i>Pseudomonas aeruginosa</i> isolates among cancer patients at the Douala Laquintinie Hospital, Littoral Region of Cameroon. Between October 2021 and March 2023, 507 study participants were recruited among whom 307 (60.55%) were cancer patients, compared to 200 (39.45%) noncancer patients. Fifty-eight <i>P. aeruginosa</i> isolates were isolated from fecal samples of forty-five cancer patients and thirteen noncancer patients using Cetrimide agar. The antimicrobial resistance profile of the isolates was determined using the Kirby-Bauer disk diffusion method. The polymerase chain reaction was used to detect the presence of extended-spectrum beta-lactamase genes among <i>P. aeruginosa</i> isolates. <i>P. aeruginosa</i> showed significant resistance rates in cancer patients compared to noncancer patients to imipenem, cefotaxime, and ceftazidime, piperacillin-tazobactam, ticarcillin-clavulanic acid, and ciprofloxacin. The multidrug resistance (MDR) rate was significantly (<i>p</i> < 0.05) higher in cancer patients than in noncancer patients. The frequency of beta-lactamase genes in the 58 ESBL-producing <i>P. aeruginosa</i> isolates was determined as 72.41% for <i>bla</i> <sub>TEM</sub>, 37.93% for <i>bla</i> <sub>OXA</sub>, 74.14% for bla<sub>CTX-M</sub>, and 44.83% for <i>bla</i> <sub>SHV</sub> genes. The study revealed an alarmingly high prevalence of fecal carriage of ESBL-producing <i>P. aeruginosa</i> with a high rate of MDR among cancer patients. It indicates that regular monitoring and surveillance of ESBL-producing <i>P. aeruginosa</i> among cancer patients are needed to improve the management of patients.</p>","PeriodicalId":14098,"journal":{"name":"International Journal of Microbiology","volume":"2024 ","pages":"7685878"},"PeriodicalIF":2.8,"publicationDate":"2024-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11222006/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141497955","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-10eCollection Date: 2024-01-01DOI: 10.1155/2024/2148676
Chandana Kumari V B, Sujay Huligere, Jayanthi M K, Khang Wen Goh, Sudhanva M Desai, Kalabharthi H L, Ramith Ramu
Boza, a cereal-based beverage popular in southeast Europe, is fortified with probiotics and is believed to positively impact the composition of the gut microflora. This investigation focused on fermented cereal-based beverage boza to identify strains of probiotic Lactobacillus spp. capable of inhibiting carbohydrate-hydrolysing enzymes α-glucosidase (AG) and α-amylase (AA). The isolated bacterial strains underwent a comprehensive assessment, including biochemical, molecular, and probiotic trait analyses such as tolerance survivability, adhesion, safety, and health-promoting attributes. We evaluated the inhibitory potential of the supernatant, cell lysate, and intact cells of Lactobacillus spp. Molecular analysis has revealed that isolates RAMULAB30 and RAMULAB29 exhibit a significant genetic similarity (>97%) to Lacticaseibacillus paracasei and Limosilactobacillus fermentum, respectively. These findings are documented in the NCBI database. They exhibited significant resistance to gastrointestinal and intestinal fluids, also indicating their potential for adhesion. Additionally, the isolates showed a significant antibacterial activity, particularly against Micrococcus luteus. They showed resistance to vancomycin and methicillin antibiotics but were more susceptible to streptomycin and ampicillin. Furthermore, the strains demonstrated antioxidant properties. To ensure their safety, a haemolytic assay was conducted despite their general recognition as safe (GRAS) status. The study primarily aimed to evaluate the inhibitory effects of the extract on enzymes AG and AA. Bacterial isolates demonstrated a significant inhibitory activity against both enzyme AG (32%-67% inhibition) and enzyme AA (18%-46% inhibition) in different forms, including supernatant (CS), lysed extract (CE), and intact cell (IC). These findings underscore the potential of bacterial isolates to inhibit the enzyme activity effectively. Furthermore, the L. fermentum RAMULAB29 and L. paracasei RAMULAB30 strains exhibit remarkable antidiabetic potential. Food products incorporating these strains have promising prospects as nutraceuticals, providing improved health benefits.
boza是一种流行于欧洲东南部的谷物饮料,添加了益生菌,被认为能对肠道微生物菌群的组成产生积极影响。这项调查的重点是发酵谷物饮料 Boza,目的是找出能够抑制碳水化合物水解酶 α-葡萄糖苷酶(AG)和 α-淀粉酶(AA)的益生菌乳酸杆菌菌株。对分离出的细菌菌株进行了全面评估,包括生化、分子和益生菌性状分析,如耐受存活性、粘附性、安全性和促进健康的属性。分子分析表明,分离物 RAMULAB30 和 RAMULAB29 分别与副酸乳杆菌(Lacticaseibacillus paracasei)和发酵乳杆菌(Limosilactobacillus fermentum)具有显著的遗传相似性(>97%)。NCBI 数据库记录了这些发现。它们对胃肠液和肠液具有明显的抵抗力,这也表明它们具有粘附潜力。此外,这些分离物还表现出了显著的抗菌活性,尤其是对黄体微球菌的抗菌活性。它们对万古霉素和甲氧西林抗生素表现出耐药性,但对链霉素和氨苄西林更敏感。此外,这些菌株还具有抗氧化特性。为了确保它们的安全性,尽管它们被普遍认为是安全的(GRAS),但还是进行了溶血试验。研究的主要目的是评估提取物对 AG 和 AA 酶的抑制作用。细菌分离物以不同形式(包括上清液(CS)、裂解提取物(CE)和完整细胞(IC))对酶 AG(抑制率为 32%-67%)和酶 AA(抑制率为 18%-46%)均表现出明显的抑制活性。这些发现强调了细菌分离物有效抑制酶活性的潜力。此外,L. fermentum RAMULAB29 和 L. paracasei RAMULAB30 菌株表现出显著的抗糖尿病潜力。含有这些菌株的食品作为营养保健品具有广阔的前景,可改善健康状况。
{"title":"Characterization of <i>Lactobacillus</i> spp. as Probiotic and Antidiabetic Potential Isolated from Boza, Traditional Fermented Beverage in Turkey.","authors":"Chandana Kumari V B, Sujay Huligere, Jayanthi M K, Khang Wen Goh, Sudhanva M Desai, Kalabharthi H L, Ramith Ramu","doi":"10.1155/2024/2148676","DOIUrl":"10.1155/2024/2148676","url":null,"abstract":"<p><p>Boza, a cereal-based beverage popular in southeast Europe, is fortified with probiotics and is believed to positively impact the composition of the gut microflora. This investigation focused on fermented cereal-based beverage boza to identify strains of probiotic <i>Lactobacillus</i> spp. capable of inhibiting carbohydrate-hydrolysing enzymes <i>α</i>-glucosidase (AG) and <i>α</i>-amylase (AA). The isolated bacterial strains underwent a comprehensive assessment, including biochemical, molecular, and probiotic trait analyses such as tolerance survivability, adhesion, safety, and health-promoting attributes. We evaluated the inhibitory potential of the supernatant, cell lysate, and intact cells of <i>Lactobacillus</i> spp. Molecular analysis has revealed that isolates RAMULAB30 and RAMULAB29 exhibit a significant genetic similarity (>97%) to <i>Lacticaseibacillus paracasei</i> and <i>Limosilactobacillus fermentum</i>, respectively. These findings are documented in the NCBI database. They exhibited significant resistance to gastrointestinal and intestinal fluids, also indicating their potential for adhesion. Additionally, the isolates showed a significant antibacterial activity, particularly against <i>Micrococcus luteus</i>. They showed resistance to vancomycin and methicillin antibiotics but were more susceptible to streptomycin and ampicillin. Furthermore, the strains demonstrated antioxidant properties. To ensure their safety, a haemolytic assay was conducted despite their general recognition as safe (GRAS) status. The study primarily aimed to evaluate the inhibitory effects of the extract on enzymes AG and AA. Bacterial isolates demonstrated a significant inhibitory activity against both enzyme AG (32%-67% inhibition) and enzyme AA (18%-46% inhibition) in different forms, including supernatant (CS), lysed extract (CE), and intact cell (IC). These findings underscore the potential of bacterial isolates to inhibit the enzyme activity effectively. Furthermore, the <i>L. fermentum</i> RAMULAB29 and <i>L. paracasei</i> RAMULAB30 strains exhibit remarkable antidiabetic potential. Food products incorporating these strains have promising prospects as nutraceuticals, providing improved health benefits.</p>","PeriodicalId":14098,"journal":{"name":"International Journal of Microbiology","volume":"2024 ","pages":"2148676"},"PeriodicalIF":2.8,"publicationDate":"2024-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11221989/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141497954","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Poultry's digestive tract lacks hydrolytic phytase enzymes, which results in chelation of dietary minerals, vital amino acids, proteins, and carbohydrates, phytate-phosphate unavailability, and contamination of the environment due to phosphorus. Therefore, it is necessary to use exogenous microbial phytases as feed additive to chicken feed to catalyze the hydrolysis of dietary phytate. Potential sources of microbial isolates that produce desired phytases for chicken feed supplementation have been isolated from agricultural croplands. It is achievable to isolate phytase-producing bacteria isolates using both broth and agar phytase screening media. Potential substrates for submerged fermentation (SmF) for bacterial phytase production and solid-state fermentation (SSF) for fungal phytase production include rice and wheat bran. Following fermentation, saturated ammonium sulphate precipitation is typically used to partially purify microbial culture filtrate. The precipitate is then desalted. Measurements of the pH optimum and stability, temperature optimum and stability, metal ions stability, specificity and affinity to target substrate, proteolysis resistance, storage stability, and in vitro feed dephosphorylation are used to perform an enzymatic evaluation of phytase as an additive for poultry feed. The growth of the feed phytase market is primarily due to the expansion of chicken farms to meet the demand for meat and eggs from humans. The Food and Drug Administration in the USA and the European Food and Safety Authority are primarily in charge of putting rules pertaining to feed phytase use in chicken feed into effect. Conclusively, important components of the production of phytase additives for poultry feed include identifying a reliable source for potential microbe isolation, selecting an economical method of phytase production, thoroughly characterizing the biochemical properties of phytase, and comprehending the size and regulation of the current feed phytase market.
{"title":"Review on Desirable Microbial Phytases as a Poultry Feed Additive: Their Sources, Production, Enzymatic Evaluation, Market Size, and Regulation.","authors":"Olyad Erba Urgessa, Rufael Koyamo, Hunduma Dinka, Ketema Tefese, Mesfin Tafesse Gemeda","doi":"10.1155/2024/9400374","DOIUrl":"10.1155/2024/9400374","url":null,"abstract":"<p><p>Poultry's digestive tract lacks hydrolytic phytase enzymes, which results in chelation of dietary minerals, vital amino acids, proteins, and carbohydrates, phytate-phosphate unavailability, and contamination of the environment due to phosphorus. Therefore, it is necessary to use exogenous microbial phytases as feed additive to chicken feed to catalyze the hydrolysis of dietary phytate. Potential sources of microbial isolates that produce desired phytases for chicken feed supplementation have been isolated from agricultural croplands. It is achievable to isolate phytase-producing bacteria isolates using both broth and agar phytase screening media. Potential substrates for submerged fermentation (SmF) for bacterial phytase production and solid-state fermentation (SSF) for fungal phytase production include rice and wheat bran. Following fermentation, saturated ammonium sulphate precipitation is typically used to partially purify microbial culture filtrate. The precipitate is then desalted. Measurements of the pH optimum and stability, temperature optimum and stability, metal ions stability, specificity and affinity to target substrate, proteolysis resistance, storage stability, and in vitro feed dephosphorylation are used to perform an enzymatic evaluation of phytase as an additive for poultry feed. The growth of the feed phytase market is primarily due to the expansion of chicken farms to meet the demand for meat and eggs from humans. The Food and Drug Administration in the USA and the European Food and Safety Authority are primarily in charge of putting rules pertaining to feed phytase use in chicken feed into effect. Conclusively, important components of the production of phytase additives for poultry feed include identifying a reliable source for potential microbe isolation, selecting an economical method of phytase production, thoroughly characterizing the biochemical properties of phytase, and comprehending the size and regulation of the current feed phytase market.</p>","PeriodicalId":14098,"journal":{"name":"International Journal of Microbiology","volume":"2024 ","pages":"9400374"},"PeriodicalIF":2.8,"publicationDate":"2024-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11221984/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141497956","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-05-29eCollection Date: 2024-01-01DOI: 10.1155/2024/6660423
Mohammed Al Qutaibi, Suresh R Kagne
Mushrooms are a valuable source of food and medicine that have been used for centuries in various cultures. They contain a variety of phytochemicals, such as terpenoids and polysaccharides, that exhibit diverse biological activities, such as antioxidant, anti-inflammatory, anticancer, antimicrobial, immunomodulatory, and antidiabetic effects. However, mushroom's phytochemical composition and bioactivity vary depending on their species, cultivation conditions, processing methods, and extraction techniques. Therefore, using reliable analytical methods and standardized protocols is important for systematically evaluating the quality and quantity of mushroom phytochemicals and their therapeutic potential. This review provides a bibliometric analysis of the recent literature on biological activities, highlights trends in the field, and highlights the countries and journals with the highest contribution. It also discusses the nutritional value of the total content of phenolic and other phytochemicals in some species of mushrooms.
{"title":"Exploring the Phytochemical Compositions, Antioxidant Activity, and Nutritional Potentials of Edible and Medicinal Mushrooms.","authors":"Mohammed Al Qutaibi, Suresh R Kagne","doi":"10.1155/2024/6660423","DOIUrl":"10.1155/2024/6660423","url":null,"abstract":"<p><p>Mushrooms are a valuable source of food and medicine that have been used for centuries in various cultures. They contain a variety of phytochemicals, such as terpenoids and polysaccharides, that exhibit diverse biological activities, such as antioxidant, anti-inflammatory, anticancer, antimicrobial, immunomodulatory, and antidiabetic effects. However, mushroom's phytochemical composition and bioactivity vary depending on their species, cultivation conditions, processing methods, and extraction techniques. Therefore, using reliable analytical methods and standardized protocols is important for systematically evaluating the quality and quantity of mushroom phytochemicals and their therapeutic potential. This review provides a bibliometric analysis of the recent literature on biological activities, highlights trends in the field, and highlights the countries and journals with the highest contribution. It also discusses the nutritional value of the total content of phenolic and other phytochemicals in some species of mushrooms.</p>","PeriodicalId":14098,"journal":{"name":"International Journal of Microbiology","volume":"2024 ","pages":"6660423"},"PeriodicalIF":3.4,"publicationDate":"2024-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11152763/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141263699","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-05-25eCollection Date: 2024-01-01DOI: 10.1155/2024/8917136
Omar Zmerli, Sara Bellali, Gabriel Haddad, Rim Iwaza, Akiko Hisada, Erino Matsumoto, Yusuke Ominami, Didier Raoult, Jacques Bou Khalil
Background: Colistin (Polymyxin E) has reemerged in the treatment of MDR Gram-negative infections. Traditional Colistin AST methods have long turnaround times and are cumbersome for routine use. We present a SEM-AST technique enabling rapid detection of Colistin resistance through direct observation of morphological and quantitative changes in bacteria exposed to Colistin.
Methods: Forty-four Gram-negative reference organisms were chosen based on their Colistin susceptibility profiles. Bacterial suspensions of ∼107 CFU/mL were exposed to Colistin at EUCAST-ECOFF, with controls not exposed, incubated at 37°C, and then sampled at 0, 15, 30, 60, and 120 minutes. Phosphotungstic Acid (PTA) staining was applied, followed by SEM imaging using Hitachi TM4000PlusII-Tabletop-SEM at ×2000, ×5000 and ×7000 magnifications. Bacterial viability analysis was performed for all conditions by quantifying viable and dead organisms based on PTA-staining and morphologic changes.
Results: We identified a significant drop in the percentage of viable organisms starting 30 minutes after exposure in susceptible strains, as compared to nonsignificant changes in resistant strains across all tested organisms. The killing effect of Colistin was best observed after 120 minutes of incubation with the antibiotic, with significant changes in morphologic features, including bacterial inflation, fusion, and lysis, observed as early as 30 minutes. Our observation matched the results of the gold standard-based broth microdilution method.
Conclusions: We provide an extended application of the proof of concept for the utilization of the SEM-AST assay for Colistin for a number of clinically relevant bacterial species, providing a rapid and reliable susceptibility profile for a critical antibiotic.
{"title":"Antimicrobial Susceptibility Testing for Colistin: Extended Application of Novel Quantitative and Morphologic Assay Using Scanning Electron Microscopy.","authors":"Omar Zmerli, Sara Bellali, Gabriel Haddad, Rim Iwaza, Akiko Hisada, Erino Matsumoto, Yusuke Ominami, Didier Raoult, Jacques Bou Khalil","doi":"10.1155/2024/8917136","DOIUrl":"10.1155/2024/8917136","url":null,"abstract":"<p><strong>Background: </strong>Colistin (Polymyxin E) has reemerged in the treatment of MDR Gram-negative infections. Traditional Colistin AST methods have long turnaround times and are cumbersome for routine use. We present a SEM-AST technique enabling rapid detection of Colistin resistance through direct observation of morphological and quantitative changes in bacteria exposed to Colistin.</p><p><strong>Methods: </strong>Forty-four Gram-negative reference organisms were chosen based on their Colistin susceptibility profiles. Bacterial suspensions of ∼10<sup>7</sup> CFU/mL were exposed to Colistin at EUCAST-ECOFF, with controls not exposed, incubated at 37°C, and then sampled at 0, 15, 30, 60, and 120 minutes. Phosphotungstic Acid (PTA) staining was applied, followed by SEM imaging using Hitachi TM4000PlusII-Tabletop-SEM at ×2000, ×5000 and ×7000 magnifications. Bacterial viability analysis was performed for all conditions by quantifying viable and dead organisms based on PTA-staining and morphologic changes.</p><p><strong>Results: </strong>We identified a significant drop in the percentage of viable organisms starting 30 minutes after exposure in susceptible strains, as compared to nonsignificant changes in resistant strains across all tested organisms. The killing effect of Colistin was best observed after 120 minutes of incubation with the antibiotic, with significant changes in morphologic features, including bacterial inflation, fusion, and lysis, observed as early as 30 minutes. Our observation matched the results of the gold standard-based broth microdilution method.</p><p><strong>Conclusions: </strong>We provide an extended application of the proof of concept for the utilization of the SEM-AST assay for Colistin for a number of clinically relevant bacterial species, providing a rapid and reliable susceptibility profile for a critical antibiotic.</p>","PeriodicalId":14098,"journal":{"name":"International Journal of Microbiology","volume":"2024 ","pages":"8917136"},"PeriodicalIF":3.4,"publicationDate":"2024-05-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11144066/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141199264","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}