{"title":"Profiling of Pharmaceutical Sludge Microbiome, Resistome and Secondary Metabolites Using Metagenomics","authors":"Shailendra Yadav, Atya Kapley","doi":"10.1007/s12088-024-01349-9","DOIUrl":"https://doi.org/10.1007/s12088-024-01349-9","url":null,"abstract":"<p>Antibiotic manufacturing sites act as the hotspot for the dissemination of antibiotic resistance. The present study explores the resistome and secondary metabolites profile associated with the sludge of two pharmaceutical industries located in Delhi and Mysore, India. It confirmed that the pharma sludge contains complex microbiome and resistome. The presence of antibiotic-resistant genes (ARGs), class I integrons, and secondary metabolite genes (NRPs and PKs) was confirmed using PCR. Metagenomic tools like ResFinder 3.2, DeepARG, BusyBee, MG-RAST, and CoMET universe were used to analyze pharmaceutical sludge metagenome. Proteobacteria was found abundant in both metagenomes, followed by firmicutes & bacteriodetes. KEGG analysis predicted the expression of vancomycin, Pandrug, antifolate resistance pathway in both metagenomes. Resfinder predicted the presence of aminoglycoside, macrolide, and sulphonamide resistance genes in both metagenomes. DeepARG analysis classified ARGs in 28 (Arbro) & 27 (Mysore) ARG category and 431 & 368 ARG class. Further, CoMET universe indicated the presence of biosynthetic gene clusters like type II polyketide biosynthesis, Nonribosomal polypeptide biosynthesis, vancomycin & tetracycline, and macrolide biosynthesis. The present study provides primary insight about the diversity of secondary metabolites clusters present in pharmaceutical sludge. Microbes residing in such environment grows under higher selection pressure and produce various secondary metabolites. These metabolites could be exploited for the discovery of novel metabolites with antimicrobial potential and combating AMR. In future, the author aims to clone metagenome in expression vector (BAC/YAC vectors) for the discovery of novel secondary metabolites.</p>","PeriodicalId":13316,"journal":{"name":"Indian Journal of Microbiology","volume":"29 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142220319","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Histo-Pathological and Serological Investigation on the Occurrence of Subclinical Paratuberculosis in Slaughtered Buffaloes in Mhow, Madhya Pradesh, India","authors":"Vinod Kumar Singh, Gaya Prasad Jatav, Renu Singh, Neeraj Gangwar, Chayanika Das, Sharad Kumar Yadav","doi":"10.1007/s12088-024-01360-0","DOIUrl":"https://doi.org/10.1007/s12088-024-01360-0","url":null,"abstract":"<p>Paratuberculosis, caused by <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> (MAP), is an incurable chronic disease affecting virtually all ruminants. Although buffaloes are usually considered to be more resistant than cattle, bubaline paratuberculosis displays similar gross and histological lesions. The association of MAP with human Crohn’s disease poses a high risk of infection for slaughterhouse and dairy farm workers. The objective of the present study was to identify subclinical paratuberculosis in buffaloes slaughtered at Mhow cantonment abattoir, Madhya Pradesh, India. A total of 150 paired samples comprising serum (N = 50), tissues of the intestine (N = 50), and mesenteric lymph nodes (N = 50) from each animal were collected randomly from 50 buffaloes over a period of 3 months to perform the diagnosis by competitive ELISA for anti-MAP IgG detection in serum samples. Tissue samples were subjected to histopathology to classify the lesions in hematoxylin–eosin-stained tissue sections and to detect acid-fast bacilli in sister tissue sections on Ziehl–Neelsen staining. Anti-MAP antibodies were detected in 22.0% (11/50) of the serum samples. Among tissue samples, grade I, II, III, and IV lesions were observed in 24, 22, 4, 0 ileum, and 20, 24, 5, and 1 mesenteric lymph node samples, respectively. Acid-fast bacilli were observed in 6 ileum and 11 mesenteric lymph node samples. Overall, histological findings were more defined and illustrative than ELISA and can be used for the identification of paratuberculosis among slaughtered animals, especially in laboratories with limited resources. The findings also suggest an urgent requirement for the adoption of paratuberculosis control measures in the area. </p>","PeriodicalId":13316,"journal":{"name":"Indian Journal of Microbiology","volume":"3 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142220315","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"HIV Cure: How Far We Have Come?","authors":"Shesh Prakash Maurya, Abhishek Shrivastav, Vivek Singh Rawat, Hitender Gautam, Bimal Kumar Das","doi":"10.1007/s12088-024-01353-z","DOIUrl":"https://doi.org/10.1007/s12088-024-01353-z","url":null,"abstract":"<p>Human Immunodeficiency Virus (HIV) is a major global healthcare burden. Current lifelong antiretroviral therapy drastically improves life expectancy but do not cure HIV. Therefore, at the existing growth rates, it is estimated that around 42 million people will be living with HIV by 2030 worldwide. A cure for HIV is need of the hour which could come in the form of remission (durable viral control without ART) or eradication (complete removal of latent replication-competent virus). In this review, we discuss recent advances in basic, applied and clinical aspects of latent HIV reservoirs including its tissue locations, cell types, cell properties, genomic integration sites and its significance, mechanism of reservoir seeding and methods to study the reservoirs. Natural models of functional cure which include elite controllers, viremic controllers, long term non-progressors and post-treatment controllers are discussed. Recent advances towards a functional HIV cure are discussed under headings; CCR5Δ32/Δ32 stem cells transplantation, shock and kill strategy, block and lock strategy, gene therapy and combined strategies.</p>","PeriodicalId":13316,"journal":{"name":"Indian Journal of Microbiology","volume":"12 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142220317","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Endophyte-Mediated Reinforcement of Morpho-Physiology of Brassica juncea Under Cadmium Toxicity","authors":"Gurvarinder Kaur, Palampreet Singh, Shruti Kaushik, Isha Madaan, Ashish Vyas, Sarvajeet Singh Gill, Geetika Sirhindi","doi":"10.1007/s12088-024-01375-7","DOIUrl":"https://doi.org/10.1007/s12088-024-01375-7","url":null,"abstract":"<p>The present study was conducted to test the efficacy of <i>Serendipita indica</i> in <i>Brassica juncea</i> against cadmium (Cd) stress. Cd is a trace element that enters into plants through contaminated soil. <i>Serendipita indica</i> is a fungal endophyte which colonizes and benefits exceptionally large group of plants. Roots of 3-days old <i>B. juncea</i> seedlings were inoculated with <i>S. indica</i> and raised in toxic Cd concentrations (10 µM, 30 µM and 50 µM Cd) till 12 days after inoculation in plant growth chamber with conditions of 16-h light/8-h dark photoperiod, temperature 25 °C (± 2 °C) and 80% relative humidity. The seedlings were tested for various morpho-physiological parameters like seed germination, plant biomass and %age heavy metal tolerance index. Also, various non-enzymatic antioxidants like vitamin A, vitamin C, phenols, flavonoids, total soluble sugars, reducing and non-reducing sugars were analyzed. Cd led to reduction in the % age germination of <i>B. juncea</i> seeds<i>. Serendipita indica</i> inoculation incremented the growth parameters like fresh weight, dry weight, root length, shoot length which were diminished by Cd stress. Enhancement of 20.66% in root length was observed in inoculated and Cd stressed seedlings. The content of phenols increased under unstressed conditions by 26.6% and by 55.29% under Cd stressed conditions in <i>S. indica</i> inoculated seedlings along with increasing contents of vitamin A and C. Overall, the content of carbohydrates was enhanced with <i>S. indica</i> inoculation. The study suggests that <i>S. indica</i> endophyte has great potential in boosting growth and physiological parameters which help in managing Cd stress in <i>B. juncea</i>.</p><h3 data-test=\"abstract-sub-heading\">Graphical Abstract</h3>","PeriodicalId":13316,"journal":{"name":"Indian Journal of Microbiology","volume":"286 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142220316","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Na He, Cunjian Yi, Qingsong Zeng, Wumei Jing, Wenrong He
{"title":"Advancements in the Vaginal Microenvironment and Regression of High-Risk Human Papillomavirus","authors":"Na He, Cunjian Yi, Qingsong Zeng, Wumei Jing, Wenrong He","doi":"10.1007/s12088-024-01369-5","DOIUrl":"https://doi.org/10.1007/s12088-024-01369-5","url":null,"abstract":"<p>Persistent infection with high-risk human papillomavirus is considered one of the main causes of cervical cancer. In recent years, there has been increasing concern about the health problems associated with HPV infection. To better understand the mechanisms by which high-risk human papillomavirus infection leads to cervical cancer, many studies have begun to explore the role of the vaginal microenvironment in this context. The female vagina is an open cavity with a special anatomical structure, which constitutes the first barrier to protect the vaginal microenvironment. At the same time, the female reproductive tract is a typical micro-ecosystem with a wide variety of microflora, which maintains a dynamic and balanced relationship of symbiosis, coexistence, and mutual control with the external environment and the host. Hormonal changes during the physiological cycle, especially estrogen, have an important influence on the structure and stability of the vaginal flora. In addition, the vagina has an important role in immune homeostasis, where the immune system plays a crucial role in maintaining tissue homeostasis, eliminating pathogens, and avoiding barrier damage. When a pathogen such as human papillomavirus enters the vagina, the immune system initiates an associated immune response to clear the virus and restore tissue health. However, in some cases, the immune system may not be able to effectively respond to human papillomavirus infection, leading to the development of persistent infections. Dysregulation of vaginal microecology may be an important factor in persistent human papillomavirus infection. This review focuses on this topic by describing the vaginal microenvironment, human papillomavirus, and the mechanisms involved in influencing the vaginal microecology causing persistent human papillomavirus infection and thus accelerating cervical carcinogenesis.</p>","PeriodicalId":13316,"journal":{"name":"Indian Journal of Microbiology","volume":"431 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142220318","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Burkholderia Cenocepacia MALDI-ToF Mass Spectra in the Assessing the Course of Lower Respiratory Tract Infection in Patients with Cystic Fibrosis","authors":"Olga Vladimirovna Kondratenko, Arina Ilinichna Sizova, Alena Anatolyevna Ereshchenko, Andrei Vladimirovich Kozlov, Danir Damirovich Ismatullin, Artem Viktorovich Lyamin, Aleksey Sergeevich Sustretov","doi":"10.1007/s12088-024-01366-8","DOIUrl":"https://doi.org/10.1007/s12088-024-01366-8","url":null,"abstract":"<p>A microbial identification method using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF) is an innovative dimension in proteomic analysis. MALDI-ToF mass spectrometry allows not only determine the species and subspecies bacteria, but also determined by proteomic analysis and the corresponding software degree of kinship analyzed strains, which allows this method to be used in epidemiological studies and in comparing strains isolated from patients with chronic infection. The aim of this study was to evaluate the possibility of using protein spectra of microorganisms obtained by MALDI-ToF mass spectrometry as additional microbiological criteria in assessing the course of the infection process caused by <i>Burkholderia cepacia</i> complex among patients with cystic fibrosis. The analysis of protein profiles, which were obtained by using MALDI-ToF mass spectrometry (Bruker Daltonik GmbH, Germany), was performed by using flexAnalisis 3.0 software (Bruker Daltonik GmbH, Germany). Differences in protein profiles of <i>Burkholderia</i> spp. isolates were found depended on the stage of infection of a cystic fibrosis patient with prolonged colonisation of the lower respiratory tract. The protein profiles of <i>Burkholderia</i> spp. isolates that formed a heterogeneous population containing both NCV and SCV morphotypes were also studied. A regular dynamic monitoring and comparison of protein profiles of microbial strains can be useful in forecasting effort of the clinical course of the disease, as well as in assessing of risks of severe infectious complications development.</p>","PeriodicalId":13316,"journal":{"name":"Indian Journal of Microbiology","volume":"6 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142220320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Carbon Nanotubes in the Diagnosis and Treatment of Ovarian Cancer","authors":"Aditi Chattaraj, Vijay Mishra, Yachana Mishra","doi":"10.1007/s12088-024-01367-7","DOIUrl":"https://doi.org/10.1007/s12088-024-01367-7","url":null,"abstract":"<p>One of the most serious gynecological diseases in the world is ovarian cancer (OC). These days, the majority of patients are identified at an advanced stage (III or IV), with subpar diagnosis resulting in a return of the illness. Conventional medicines fail as a result of issues with early illness identification and treatment processing, including issues with dosage delivery, side effects, and treatment resistance. The carbon nanotube (CNT)-based drug delivery systems for specific OC therapy are highlighted in this review. These systems have several advantages against free drugs, including nontoxicity, biological compatibility, high biodegradability, increased therapeutic impact, and non-inflammatory effects. Crucially, functionalized CNTs with particular ligands like cancer antigen (CA125), Human epididymis protein 4 (HE4), Mucin 1, and folic acid (FA) allow for selective targeting of OC and ultimately increase therapeutic potential in comparison to their nonfunctionalized counterparts. This review focused on the potential applications of CNTs in the detection and treatment of OC, as well as their present status and future clinical developments.</p><h3 data-test=\"abstract-sub-heading\">Graphical Abstract</h3>\u0000","PeriodicalId":13316,"journal":{"name":"Indian Journal of Microbiology","volume":"193 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141933424","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Microbial Biopigment Production by Rhodotorula sp. Using Horticultural Waste; Its Functionality, Stability and Applications","authors":"Shubham Mahajan, Pooja Nikhanj, Arashdeep Singh","doi":"10.1007/s12088-024-01370-y","DOIUrl":"https://doi.org/10.1007/s12088-024-01370-y","url":null,"abstract":"<p>Potato waste can serve as the growth media component for microorganism’s growth owing to its nutritional profile. The study evaluates different horticultural wastes to utilize it as media component to develop a cheaper media formulation for biopigment production using <i>Rhodotorula</i> sp. MTCC 1446. Proximate analysis of wastes revealed the optimum level of total sugars, reducing sugars, titratable acidity, macro and micronutrients in potato waste for growth of <i>Rhodotorula</i> sp. yeast culture. Concentration optimization showed the 5.0% (w/v) potato waste as media component resulted in 2.82 ± 0.8 g/kg biomass and 259.46 ± 9.5 µg/g biopigment productions. Standardization of growth conditions were carried out implementing Response Surface Methodology. Optimized growth conditions for <i>Rhodotorula</i> sp. were found to be pH 7.0, 30 °C, 66 h incubation time and 362.5 g DAHP addition that lead to production of 34.60 g/L biomass and 3345.82 µg/g biopigment. Validation at 3L scale showed the production of 32.6 g/L biomass and 3152.42 µg/g biopigment with the desirability of 98%. Characterization discovered the presence of carotenoids having antimicrobial, anticancerous, antioxidant properties and stability studies revealed the thermostable (up to 70 °C) and UV light and sunlight stability for 4 h of biopigment. Utilization of biopigment in food products increased the nutraceutical value of the product by increasing its phenolic content as well as appearance of the respected food items. In conclusion, the biopigment produced growing <i>Rhodotorula</i> sp. on potato waste based media under optimal conditions represents a viable natural substitute to artificial food colors in food industry.</p>","PeriodicalId":13316,"journal":{"name":"Indian Journal of Microbiology","volume":"22 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141933423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Innovative Biosensor Technologies in the Diagnosis of Urinary Tract Infections: A Comprehensive Literature Review","authors":"Haluk Çelik, Balım Bengisu Caf, Gizem Çebi","doi":"10.1007/s12088-024-01359-7","DOIUrl":"https://doi.org/10.1007/s12088-024-01359-7","url":null,"abstract":"<p>Urinary tract infections (UTIs) are prevalent bacterial infections globally, posing significant challenges due to their frequency, recurrence, and antibiotic resistance. This review delves into the advancements in UTI diagnostics over the past decade, particularly focusing on the development of biosensor technologies. The emergence of biosensors, including microfluidic, optical, electrochemical, immunosensors, and nanotechnology-based sensors, offers enhanced diagnostic accuracy, reduced healthcare costs. Despite these advancements, challenges such as technical limitations, the need for cross-population validation, and economic barriers for widespread implementation persist. The integration of artificial intelligence and smart devices in UTI diagnostics, highlighting the innovative approaches and their implications for patient care. The article envisions a future where multidisciplinary research and innovation overcome current obstacles, fully leveraging the potential of biosensor technologies to transform biosensor-based UTIs diagnosis. The ultimate goal is to achieve rapid, accurate, and non-invasive diagnostics, making healthcare more accessible and effective.</p>","PeriodicalId":13316,"journal":{"name":"Indian Journal of Microbiology","volume":"69 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141881330","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Co-existence of Metal Transporter Genes and Heavy Metal Bioremediation Potential of Bacterial Strains from the Effluent Water","authors":"Harpreet Kaur, Priya Katyal, Simranpreet Kaur Natt, Yogesh Vikal, Sumita Chandel","doi":"10.1007/s12088-024-01362-y","DOIUrl":"https://doi.org/10.1007/s12088-024-01362-y","url":null,"abstract":"<p>Heavy metal contamination and accumulation pose a major threat to all life forms in the environment due to its toxic nature, non-biodegradable properties and accumulative behavior. The present study aims to investigate the metals resistance mechanisms, removal capacity and bioremediation potential of the high metal-resistant indigenous bacterial species <i>Bacillus thuringiensis</i> strain BHR1, <i>Bacillus cereus</i> strain BHR2, <i>Enterobacter cloacae</i> strain BHR4, <i>Bacillus pumilus</i> strain BHR5, <i>Bacillus altitudinis</i> strain BHR6 and <i>Klebsiella pneumonia</i> strain BHR8 isolated from the effluent water. Moreover, the presence of specific metal transporter genes such as <i>chrA</i>, <i>chrB</i>, <i>pbrA</i> and <i>nccA</i> in these isolates indicated their role in metal resistance and removal. Based on minimum inhibitory concentration, bacterial isolates namely, BHR1 and BHR5 exhibited high metal tolerance against Cr (510 and 550 ppm), Pb (250 and 300 ppm), Ni (300 and 150 ppm) and As (200 and 120 ppm), respectively. The bioremediation potential of these bacterial isolates (BHR1 and BHR5) was evaluated under batch-scale conditions. Our results revealed that the addition of nutrients, as well as modification of pH to 6.0, enhances the bioremediation potential of BHR1 enabling the reduction of Pb, Cr, Ni and Zn by 60.9%, 31%, 22.7% and 22%, respectively. These findings suggested the potential application of these metal resistant bacterial isolates for the bioremediation of heavy metal contaminated effluents.</p>","PeriodicalId":13316,"journal":{"name":"Indian Journal of Microbiology","volume":"152 1","pages":""},"PeriodicalIF":3.0,"publicationDate":"2024-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141881331","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}