Foodborne pathogens and disease最新文献

{"title":"Rosalind Franklin Society Proudly Announces the 2023 Award Recipient for <i>Foodborne Pathogens and Disease</i>.","authors":"Megan E Jacob","doi":"10.1089/fpd.2024.79803.rfs2023","DOIUrl":"https://doi.org/10.1089/fpd.2024.79803.rfs2023","url":null,"abstract":"","PeriodicalId":12333,"journal":{"name":"Foodborne pathogens and disease","volume":"21 9","pages":"525"},"PeriodicalIF":1.9,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142139708","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Into the Jungle of Biological Agents of Foodborne Diseases: Time to Put Some Order for the French Risk Manager.","authors":"Frédérique Audiat-Perrin, Laurent Guillier, Jean-Christophe Augustin, Gilles Bornert, Michel Federighi, Michel Gautier, Nathalie Jourdan-da Silva, Régis Pouillot, Myriam Merad, Moez Sanaa, Pauline Kooh","doi":"10.1089/fpd.2023.0105","DOIUrl":"10.1089/fpd.2023.0105","url":null,"abstract":"<p><p>Consumers can be exposed to many foodborne biological hazards that cause diseases with varying outcomes and incidence and, therefore, represent different levels of public health burden. To help the French risk managers to rank these hazards and to prioritize food safety actions, we have developed a three-step approach. The first step was to develop a list of foodborne hazards of health concern in mainland France. From an initial list of 335 human pathogenic biological agents, the final list of \"retained hazards\" consists of 24 hazards, including 12 bacteria (including bacterial toxins and metabolites), 3 viruses and 9 parasites. The second step was to collect data to estimate the disease burden (incidence, Disability Adjusted Life Years) associated with these hazards through food during two time periods: 2008-2013 and 2014-2019. The ranks of the different hazards changed slightly according to the considered period. The third step was the ranking of hazards according to a multicriteria decision support model using the ELECTRE III method. Three ranking criteria were used, where two reflect the severity of the effects (Years of life lost and Years lost due to disability) and one reflects the likelihood (incidence) of the disease. The multicriteria decision analysis approach takes into account the preferences of the risk managers through different sets of weights and the uncertainties associated with the data. The method and the data collected allowed to estimate the health burden of foodborne biological hazards in mainland France and to define a prioritization list for the health authorities.</p>","PeriodicalId":12333,"journal":{"name":"Foodborne pathogens and disease","volume":" ","pages":"536-545"},"PeriodicalIF":1.9,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141534112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Antibacterial Effect of <i>Euryale ferox</i> Seed Shell Polyphenol Extract on <i>Salmonella Typhimurium</i>.","authors":"Meihong Cai, Qiaoqiao Xu, Shili Zhao, Xianhan Zhou, Yuelin Cai, Xingle He","doi":"10.1089/fpd.2023.0185","DOIUrl":"10.1089/fpd.2023.0185","url":null,"abstract":"<p><p>This study aimed to evaluate the effects of <i>Euryale ferox</i> Seed Shell Polyphenol Extract (EFSSPE) on a foodborne pathogenic bacterium. EFSSPE showed antimicrobial activity toward <i>Salmonella</i> Typhimurium CICC 22956; the minimum inhibitory concentration of EFSSPE was 1.25 mg/mL, the inhibition curve also reflected the inhibitory effect of EFSSPE on the growth of <i>S</i>. Typhimurium. Detection of alkaline phosphatase outside the cell revealed that EFSSPE treatment damaged the cell wall integrity of <i>S.</i> Typhimurium. EFSSPE also altered the membrane integrity, thereby causing leaching of 260-nm-absorbing material (bacterial proteins and DNA). Moreover, the activities of succinate dehydrogenase and malate dehydrogenase were inhibited by EFSSPE. The hydrophobicity and clustering ability of cells were affected by EFSSPE. Scanning electron microscopy showed that EFSSPE treatment damaged the morphology of the tested bacteria. These results indicate that EFSSPE can destroy the cell wall integrity and alter the permeability of the cell membrane of <i>S.</i> Typhimurium.</p>","PeriodicalId":12333,"journal":{"name":"Foodborne pathogens and disease","volume":" ","pages":"570-577"},"PeriodicalIF":1.9,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141491481","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Prediction of <i>Listeria monocytogenes</i> Clonal Complexes from Multilocus Variable Number Tandem Repeat Analysis Patterns Using a Machine Learning Approach.","authors":"Nicholas Andrews, Natalia Unrath, Patrick Wall, James F Buckley, Séamus Fanning","doi":"10.1089/fpd.2023.0163","DOIUrl":"10.1089/fpd.2023.0163","url":null,"abstract":"<p><p>Multilocus variable number tandem repeat analysis (MLVA) is a molecular subtyping technique that remains useful for those without the resources to access whole genome sequencing for the tracking and tracing of bacterial contaminants. Unlike techniques such as multilocus sequence typing (MLST) and pulsed-field gel electrophoresis, MLVA did not emerge as a standardized subtyping method for <i>Listeria monocytogenes</i>, and as a result, there is no reference database of virulent or food-associated MLVA subtypes as there is for MLST-based clonal complexes (CCs). Having previously shown the close congruence of a 5-loci MLVA scheme with MLST, a predictive model was created using the XGBoost machine learning (ML) technique, which enabled the prediction of CCs from MLVA patterns with ∼85% (±4%) accuracy. As well as validating the model on existing data, a straightforward update protocol was simulated for if and when previously unseen subtypes might arise. This article illustrates how ML techniques can be applied with elementary coding skills to add value to previous-generation molecular subtyping data in-built food processing environments.</p>","PeriodicalId":12333,"journal":{"name":"Foodborne pathogens and disease","volume":" ","pages":"593-599"},"PeriodicalIF":1.9,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141534113","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular Epidemiology of Carbapenemase-Producing <i>Pseudomonas aeruginosa</i>: An Iranian Referral Hospital-Based Study.","authors":"Shima Mahmoudi, Erfaneh Jafari, Babak Pourakbari, Shiva Poormohammadi, Reihaneh Hosseinpour Sadeghi, Setareh Mamishi","doi":"10.1089/fpd.2024.0066","DOIUrl":"https://doi.org/10.1089/fpd.2024.0066","url":null,"abstract":"<p><p>In recent years, there has been a significant increase in infections caused by carbapenemase-producing strains, with carbapenem-resistant <i>Pseudomonas aeruginosa</i> (CRPA) emerging as a priority pathogen according to the World Health Organization. This study aimed to evaluate the molecular epidemiology of CRPA isolated from patients referred to Children's Medical Center in Tehran, Iran. <i>P. aeruginosa</i> isolates collected from different children's wards were screened for common carbapenem-resistant genes by polymerase chain reaction (PCR). Genetic relatedness between isolates was assessed by pulsed-field gel electrophoresis (PFGE). The study included 133 participants, with 50% being male, and revealed a median age of 2 years (interquartile range: 6 months to 6 years). Carbapenem resistance was detected in 15% of cases (<i>n</i> = 20), with CRPA isolates predominantly found in the emergency ward (60%). The median age of patients with CRPA was significantly higher than those with carbapenem-susceptible <i>P. aeruginosa</i> (6 years vs. 1 year). PCR analysis revealed metallo-β-lactamase production in 45% of CRPA isolates (<i>n</i> = 9), with <i>bla_NDM</i> being the most prevalent gene. PFGE analysis of the CRPA isolates identified three clusters (Cluster I, II, and III). Cluster I, comprising 65% of all isolates (<i>n</i> = 13), was predominantly found in the emergency ward. Notably, <i>bla_NDM</i>-producing strains were prevalent in the emergency ward. Our study highlights the significant prevalence of CRPA in the emergency ward of our hospital and underscores the importance of targeted surveillance and infection control measures to curb its spread within health care settings.</p>","PeriodicalId":12333,"journal":{"name":"Foodborne pathogens and disease","volume":" ","pages":""},"PeriodicalIF":1.9,"publicationDate":"2024-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142105980","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Prevalence of <i>Brucella</i> Spp. in Dairy Products in Iran: A Systematic Review and Meta-Analysis.","authors":"Narges Shahbazpour, Elahesadat Hosseini, Seyed Ehsan Beladian Behbahan, Fatemeh Esfarjani, Seyed Amir Mohamad Mortazavian, Abdol-Samad Abedi, Slim Smaoui","doi":"10.1089/fpd.2024.0093","DOIUrl":"https://doi.org/10.1089/fpd.2024.0093","url":null,"abstract":"<p><p>Brucellosis, a significant zoonotic disease, threatens food safety substantially, particularly in developing nations such as the Middle East. This study aimed to comprehensively assess the prevalence of <i>Brucella</i> spp. in Iranian milk and dairy products through a systematic review and meta-analysis. A thorough search of international and domestic databases from January 2008 to October 2023 identified 38 relevant studies encompassing 11,130 samples for meta-analysis. The overall prevalence of <i>Brucella</i> spp. in Iranian dairy products was 22% (95% CI: 16-28%). The highest and lowest overall prevalence of <i>Brucella</i> spp. in milk were found in raw goat milk 27% (95% CI: 11-42%) and raw camel milk 15% (95% CI: -0.42 to 72%), respectively. The overall prevalence of traditional cheese, ice cream, and cream is estimated to be 9% (95% CI: -16 to 35%), 2% (95% CI: -2.78 to 2.82%), and 9% (95% CI: -0.94 to 1.12%). Geographical disparities were evident, with Zanjan province reporting the highest contamination rate, 53%, while Razavi Khorasan province had the lowest, 1%. However, the prevalence of <i>Brucella</i> spp. in Iranian dairy products has fluctuated over time, with a significant association between the study year and sample size. Comprehensive planning, robust policy implementation, and rigorous monitoring are imperative to mitigate and ultimately eliminate <i>Brucella</i> contamination in dairy products effectively. Further research is essential to refine prevalence estimates and develop targeted prevention strategies to safeguard public health.</p>","PeriodicalId":12333,"journal":{"name":"Foodborne pathogens and disease","volume":" ","pages":""},"PeriodicalIF":1.9,"publicationDate":"2024-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142105981","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Contamination Status and Serotypes Distribution of <i>Salmonella</i> in Food in Yantai City, China: A 14-Year Continuous Monitoring Study.","authors":"Fengguang Dong, Guiqiang Wang, Xueying Feng, Chunbo Gong, Wenjuan Liu, Songsong Wang, Yiyi Zhang, Yapeng Huo, Youxia Chen, Hongtao Wang","doi":"10.1089/fpd.2024.0055","DOIUrl":"https://doi.org/10.1089/fpd.2024.0055","url":null,"abstract":"<p><p><i>Salmonella</i> is a foodborne zoonotic pathogen that threatens food safety and public health. However, few people have conducted long-term and systematic studies on <i>Salmonella</i> contamination in food in Yantai City. In order to investigate the situation of <i>Salmonella</i> contamination in food and improve the ability of early warning and control of foodborne diseases, a total of 3420 samples from 20 categories were collected from 13 monitoring points in Yantai City, from 2010 to 2023. The difference in detection rate and bacterial strain of different monitoring points, different types, and different sources of samples was compared. Of the 3420 samples, 80 were positive with a detection rate of 2.34%. <i>Salmonella</i> detection rates were significantly different for samples collected at different monitoring sites. <i>Salmonella</i> was detected only in meat and meat products and catering food, and none of the other types were detected. The detection rate of <i>Salmonella</i> was higher in raw animal meat and raw poultry. Samples collected at the market stage had the highest detection rate (5.81%), and there was a significant difference in detection rate between samples from different sources (χ² = 36.93, <i>p</i> < 0.05). Eighty-one strains of <i>Salmonella</i> were detected out of 3420 samples (2 different strains were detected in 1 positive sample). The serological test identified 8 groups and 27 serotypes. The dominant serum groups were group B 30.86% (25/81), group E1 23.46% (19/81), and group D 16.05% (13/81). The main dominant serotypes were <i>Salmonella</i> <i>give</i> 17.28% (14/81), <i>Salmonella</i> <i>enteritidis</i> 16.05% (13/81), and <i>Salmonella</i> <i>derby</i> 13.58% (11/81). Meat and meat products and catering food were the main food products contaminated with <i>Salmonella</i>. The resulting secondary contamination is the hidden threat of foodborne diseases and should be given sufficient attention.</p>","PeriodicalId":12333,"journal":{"name":"Foodborne pathogens and disease","volume":" ","pages":""},"PeriodicalIF":1.9,"publicationDate":"2024-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141909835","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Duplex PCR Assay for Rapid Detection of <i>Klebsiella pneumoniae</i> and <i>Chryseobacterium</i> in Large Yellow Croaker Fish.","authors":"Gaowei Hu, Longfei Yin, Xi Luo, Yingjie Miao, Jianyun Yu","doi":"10.1089/fpd.2023.0149","DOIUrl":"10.1089/fpd.2023.0149","url":null,"abstract":"<p><p>Both <i>Klebsiella pneumoniae</i> and <i>Chryseobacterium</i> cause an increasing number of diseases in fish, resulting in great economic losses in aquaculture. In addition, the disease infected with <i>Klebsiella pneumoniae</i> or <i>Chryseobacterium</i> exhibited the similar clinical symptoms in aquatic animals. However, there is no effective means for the simultaneous detection of co-infection and discrimination them for these two pathogens. Here, we developed a duplex polymerase chain reaction (PCR) method based on the outer membrane protein A (<i>ompA</i>) gene of <i>Klebsiella pneumoniae</i> and <i>Chryseobacterium</i>. The specificity and validity of the designed primers were confirmed experimentally using simplex PCR. The expected amplicons for <i>Klebsiella pneumoniae</i> and <i>Chryseobacterium</i> had a size of 663 and 1404 bp, respectively. The optimal condition for duplex PCR were determined to encompass a primer concentration of 0.5 μM and annealing temperature of 57°C. This method was analytical specific with no amplification being observed from the genomic DNA of <i>Escherichia coli</i>, <i>Vibrio harveyi</i>, <i>Pseudomonas plecoglossicida</i>, <i>Aeromonas hydrophila</i> and <i>Acinetobacter johnsonii</i>. The limit of detection was estimated to be 20 fg of genomic DNA for <i>Chryseobacterium</i> and 200 fg for <i>Klebsiella pneumoniae</i>, or 100 colony-forming units (CFU) of bacterial cells in both cases. The duplex PCR was capable of simultaneously amplifying target fragments from genomic DNA extracted from the bacteria and fish liver. For practical validation of the method, 20 diseased fish were collected from farms, among which 4 samples were PCR-positive for <i>Klebsiella pneumoniae</i> and <i>Chryseobacterium</i>. The duplex PCR method developed here is time-saving, specific, convenient, and may prove to be an invaluable tool for molecular detection and epidemiological investigation of <i>Klebsiella pneumoniae</i> and <i>Chryseobacterium</i> in the field of aquaculture.</p>","PeriodicalId":12333,"journal":{"name":"Foodborne pathogens and disease","volume":" ","pages":"508-516"},"PeriodicalIF":1.9,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140850108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation and Biological Characteristics of a Novel Phage and Its Application to Control <i>Vibrio Parahaemolyticus</i> in Shellfish Meat.","authors":"Wenting Liu, Yiming Wu, Huajuan Wang, Hongxun Wang, Min Zhou","doi":"10.1089/fpd.2023.0133","DOIUrl":"10.1089/fpd.2023.0133","url":null,"abstract":"<p><p><i>Vibrio parahaemolyticus</i> is a common foodborne pathogenic bacterium. With the overuse of antibiotics, an increasing proportion of drug-resistant strains are emerging, which puts enormous pressure on public health. In this study, a <i>V. parahaemolyticus</i>-specific phage, VP41s3, was isolated. The head length, width, and tail length of the phage were 77.7 nm, 72.2 nm, and 17.5 nm, respectively. It remained active in the temperature range of 30-50°C and pH range of 4-11. The lytic curve of phage VP41s3 showed that the host bacteria did not grow until 11 h under phage treatment at MOI of 1000, indicating that the phage had good bacteriostatic ability. When it was added to shellfish contaminated with <i>V. parahaemolyticus</i> (15°C, 48 h), the number of bacteria in the experimental group was 2.11 log₁₀ CFU/mL lower than that in the control group at 24 h. Furthermore, genomic characterization and phylogenetic analysis indicated that phage VP41s3 was a new member of the <i>Podoviridae</i> family. The genome contained 50 open reading frames (ORFs), in which the ORF19 (thymidine kinase) was an enzyme involved in the pyrimidine salvage pathway, which might lead to the accelerated DNA synthesis efficiency after phage entered into host cells. This study not only contributed to the improvement of phage database and the development of beneficial phage resources but also revealed the potential application of phage VP41s3 in food hygiene and safety.</p>","PeriodicalId":12333,"journal":{"name":"Foodborne pathogens and disease","volume":" ","pages":"467-477"},"PeriodicalIF":1.9,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140957168","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Induction of <i>Salmonella</i> Enteritidis into a Viable but Nonculturable State by Cinnamaldehyde and Its Resuscitation.","authors":"Ziying Lu, Tong Xing, Zepeng Zhao, Zefeng Li, Dongping Hou, Zhuolin Ma, Siyi Chen, Yuheng Yang, Shaoting Li, Hongmei Zhang","doi":"10.1089/fpd.2023.0186","DOIUrl":"10.1089/fpd.2023.0186","url":null,"abstract":"<p><p>Trans-cinnamaldehyde (TC), a typical plant-derived compound, has been widely used in the control of foodborne pathogen contamination. Nevertheless, the risk associated with the occurrence of viable but nonculturable (VBNC) bacteria induced by TC remains unclear. The results of this study showed that <i>Salmonella</i> Enteritidis (<i>S.</i> Enteritidis) entered the VBNC state after being induced by TC at a minimum inhibitory concentration of 312.5 μg/mL and survived for at least 22 days under TC treatment. Enhanced resistance was found against heat treatment (75°C, 30 s), antibiotics (i.e., ampicillin, ceftriaxone sodium, chloramphenicol), and hydrogen peroxide (3%) in VBNC <i>S.</i> Enteritidis. A synergistic effect against VBNC <i>S.</i> Enteritidis occurred when TC was combined with acid treatment, including lactic acid and acetic acid (pH = 3.5). VBNC and resuscitated <i>S</i>. Enteritidis by sodium pyruvate treatment (100 mM) were found to retain the infectious ability to Caco-2 cells. Relative expression levels of the stress-related genes <i>relA</i>, <i>spoT</i>, <i>ppx</i>, <i>lon</i>, <i>katG</i>, <i>sodA</i>, <i>dnaK</i>, and <i>grpE</i> were upregulated in VBNC <i>S</i>. Enteritidis. Accumulation of reactive oxygen species (ROS) and protein aggregates was observed in VBNC cells. Besides, the resuscitation of VBNC cells was accompanied with clearance of ROS and protein aggregates. In summary, this study presents a comprehensive characterization of stress tolerance and resuscitation of VBNC <i>S</i>. Enteritidis induced by cinnamaldehyde, and the results provide useful information for the development of effective control strategy against VBNC pathogenic bacteria in food production.</p>","PeriodicalId":12333,"journal":{"name":"Foodborne pathogens and disease","volume":" ","pages":"499-507"},"PeriodicalIF":1.9,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140853382","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}