Whole-Genome Analysis of blaNDM-Bearing Proteus mirabilis Isolates and mcr-1-Positive Escherichia coli Isolates Carrying blaNDM from the Same Fresh Vegetables in China.

The global spread of colistin or carbapenem-resistant Enterobacteriaceae (CRE) has been a pressing threat to public health. Members of Enterobacteriaceae, especially Proteus mirabilis and Escherichia coli, have been prevalent foodborne pathogens and such pathogens from fresh vegetables have triggered foodborne illness in China. However, reports about CRE, especially P. mirabilis from fresh vegetables, are still lacking. In this study, we identified five blaNDM-positive P. mirabilis and five blaNDM-positive generic E. coli concurrently from five fresh vegetables in two markets from China, and four of the five E. coli also carried mcr-1. The 10 isolates were characterized with methods including antimicrobial susceptibility testing, conjugation, whole-genome sequencing and phylogenetic analysis. All 10 isolates were multidrug-resistant (MDR). blaNDM-5 in five E. coli isolates and one P. mirabilis carrying blaNDM-5 was located on similarly transferable IncX3 plasmids, while transferably untypable plasmids were the carriers of blaNDM-1 in four P. mirabilis isolates from different types of vegetables/markets. mcr-1 in the four blaNDM-5-positive E. coli was located on similarly non-conjugative IncHI2 MDR plasmids lacking transfer region. Notably, ISCR1 complex class 1 integron capable of capturing blaNDM-1 was found on all untypable plasmids from P. mirabilis, and five copies of ISCR1 complex class 1 integron containing blaNDM-1 even occurred in one P. mirabilis, which showed high-level carbapenem resistance. Plasmid and phylogenetic analysis revealed that the blaNDM-positive P. mirabilis and E. coli from fresh vegetables might be derived from animals and transmitted to humans via the food chain. The concurrence of blaNDM-positive P. mirabilis and E. coli carrying both mcr-1 and blaNDM in different types of fresh vegetables eaten raw is alarming and threatens food safety. Sustained surveillance of these foodborne pathogens among fresh vegetables is urgent to ensure the health of food consumers. We report for the first time the concurrence of blaNDM-positive P. mirabilis and mcr-1-bearing E. coli carrying blaNDM from the same fresh vegetables.

Bibliometric analysis of research trends in relationship between sarcopenia and surgery.

Background: The relationship between sarcopenia and surgery has attracted an increasing number of researchers in recent years. Our study aimed to identify the current research hotspot and status in this field by using bibliometric and visualization analysis. Methods: Publications about the relationship between sarcopenia and surgery that met the inclusion criteria were collected from the Science Citation Index Expanded. The bibliometric and visualized studies were performed using VOSviewer, and R. Results: A total of 2,261 documents on the relationship between sarcopenia and surgery were included in our study. These articles were written by 13,757 authors from 2,703 institutions in 70 countries and were published in 772 journals. The USA is the most productive and influential country in this field (524 publications and 15,220 citations). The Udice French Research Universities was the most productive institution in this field (57 publications), and the University of Alberta had the largest number of citations. Annuals of Surgical Oncology published the most studies in this field. Shen Xian was the most productive author in this field (number of publications = 19), and Baracos Vickie was the most influential author, whose studies in this field had been cited 2,209 times. The cluster analysis was performed and visualized, and the keywords were classified into 6 clusters: Cluster 1 (body composition and nutrition), Cluster 2 (sarcopenia), Cluster 3 (malnutrition and cachexia), Cluster 4 (cancer surgery), Cluster 5 (elderly and frailty), Cluster 6 (neuromuscular scoliosis). Conclusion: The relationship between sarcopenia and surgery was still a controversial and well-discussed topic in recent years. Our study showed that the study in this field mainly focused on sarcopenia, oncology surgery, orthopedics, and nutrition.

Emergence of two Escherichia coli strains co-harboring mcr-1 and bla NDM in fresh vegetables from China.

BACKGROUND: The concurrence of mcr and carbapenemase genes among Enterobacteriaceae has been a great clinical concern. In our study, we aimed to investigate the prevalence of mcr-positive carbapenem-resistant Enterobacteriaceae (CRE) in fresh vegetables and shed light on the possibility of transmission of mcr-positive CRE via fresh vegetables. METHODS: In this study, 712 fresh vegetable samples from 10 provinces in China were collected between May 2017 and Dec 2018 and were screened for mcr and carbapenemase genes. Antibiotic susceptibilities for isolates co-harboring carbapenemase genes and mcr were determined by an agar dilution or a broth microdilution method. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) analysis were also performed. Transferability of the carbapenemase/mcr-bearing plasmids was determined by conjugation, replicon typing and S1-PFGE-Southern blotting. The sequences of these plasmids were analyzed by using whole-genome sequencing with Illumina Hiseq platform. RESULTS: Two E. coli isolates concomitantly carrying mcr-1 and bla NDM-5/9 from leaf rape and spinach, respectively, were found and both isolates showed multidrug resistance. Notably, mcr-1-positive 690 harboring bla NDM-5 and 701 carrying bla NDM-9 belonged to ST156 and ST2847, respectively, similar to the prevalent MLST types of E. coli co-carrying mcr-1 and bla NDM from avian in our previous study. mcr-1 was on ~33-kb IncX4 plasmid or ~60-kb IncI2 plasmid, while bla NDM-5/9 was on ~46-kb IncX3 plasmid or ~120-kb untypable plasmid. The plasmids were highly similar to those from animals and clinical patients reported in various countries.Conclusion: E. coli isolates concomitantly carrying mcr-1 and bla NDM-5/9 in fresh vegetables may serve as a direct source of pathogens in humans, and such discovery in fresh vegetables emphasizes the importance of prompt surveillance and intervention in limiting the spread of E. coli co-carrying bla NDM and mcr-1. To our knowledge, this is the first report of Enterobacteriaceae co-carrying bla NDM and mcr-1 in fresh vegetables.

Colistin-Resistant mcr-Positive Enterobacteriaceae in Fresh Vegetables, an Increasing Infectious Threat in China.

The presence of mobilized colistin resistance (mcr) genes is a global concern. However, data concerning mcr in fresh vegetables, a reservoir for antibiotic resistance genes, are still rare. In this study, mcr genes were analysed in 528 vegetable samples from 53 supermarkets or farmer's markets in 23 cities of 9 provinces in China, and the mcr-positive Enterobacteriaceae were characterized. Nineteen (3.6%) samples carried one or more mcr-positive isolates, and the highest three detection rates were found in carrot, pak choi and green pepper. Twenty-four mcr-1-positive isolates (23 Escherichia coli and one Enterobacter cloacae) were obtained, and E. coli isolates showed high genetic diversity. Different multilocus sequence type (MLST) isolates were also observed within the same sample. All 24 isolates showed multidrug resistance, and 14 carried blaCTX-M genes. Most isolates harbored similarly conjugative IncX4-type (∼33 kb) or IncI2-type (∼60 kb) mcr-1-bearing plasmids. The sequenced prevalent IncX4 plasmid and IncI2 plasmid from tomato were similar to the relevant plasmids from animals and clinical isolates in various countries. mcr-1-bearing IncHI2/ST3 plasmid highly similar to that carrying 14 resistance genes from E. coli of chicken was also observed. In conclusion, a high prevalence of mcr-1 in fresh vegetables was found in China, and the dissemination of mcr-1 was mediated by similar IncX4 or IncI2 plasmids. The plasmids from vegetables showed high similarity to plasmids from clinical isolates, indicating MCR-1-producers in ready-to-eat vegetables may pose a huge threat to public health and measures need to be taken to ensure food safety.

Characterization of Highly Prevalent Plasmids Coharboring mcr-1, oqxAB, and blaCTX-M and Plasmids Harboring oqxAB and blaCTX-M in Escherichia coli Isolates from Food-Producing Animals in China.

The emergence and spread of multidrug resistance (MDR) plasmids carrying the colistin resistance gene mcr-1 has become a major public health concern. However, there is a paucity of data regarding the prevalence of mcr-1 plasmids concomitantly carrying blaCTX-M and oqxAB, an efflux pump that confers resistance to multiple agents. In this study, we determined the prevalence and characteristics of plasmids coharboring mcr-1, oqxAB, and blaCTX-M as well as those harboring oqxAB and blaCTX-M in Escherichia coli from food-producing animals. We isolated 493 E. coli strains, and mcr-1, blaNDM, and blaCTX-M were present in 140 (28.4%), 51 (10.3%), and 195 (39.6%) of the isolates, respectively. The two most prevalent plasmid-mediated quinolone resistance genes were oqxAB (34.5%) and qnrS (29.4%). Nine IncHI2/ST3 plasmids co-carrying mcr-1, oqxAB, and blaCTX-M were found, and similar IncHI2/ST3 plasmids mediated dissemination of these resistance genes. Two sequenced MDR IncHI2/ST3 plasmids coharboring mcr-1, oqxAB, and blaCTX-M showed high similarity to reference plasmid pHNSHP45-2, although they were from different regions in China. Colocalization of oqxAB and blaCTX-M on the same plasmid was found in 28 isolates, including the nine plasmids harboring mcr-1. The co-dissemination of oqxAB and blaCTX-M was mediated by diverse F33:A-:B- plasmids and similar IncHI2/ST3 plasmids. Pulsed-field gel electrophoresis and multilocus sequence typing analysis of donor isolates revealed heterogeneous patterns indicating that clonal dissemination was unlikely. The high incidence of similar IncHI2/ST3 plasmids simultaneously possessing mcr-1, oqxAB, and blaCTX-M poses a great threat to public health.

Characterization and Molecular Mapping of Two Novel Genes Resistant to Pythium Stalk Rot in Maize.

Pythium stalk rot caused by Pythium inflatum is becoming a more and more serious disease in maize, and it has caused severe yield loss in China in recent years. Deployment of resistant maize varieties is the most effective way to control this disease. Searching for the resistant maize germplasm and identifying the resistance genes are the vital processes in the breeding program. The maize inbred line X178 previously showed high resistance to Pythium stalk rot. Thus, this study aimed to reveal the gene(s) resistance to Pythium stalk rot in X178 by resistance inheritance analysis using the derived F2 and F2:3 genetic populations. The results showed that two independently inherited dominant genes, designated RpiX178-1 and RpiX178-2, carried by X178 are responsible for its resistance relative to the susceptible parent Ye107; they are located on regions of maize chromosome (chr.) 1 bin 1.09 and chr. 4 bin 4.08, respectively, and flanked by markers umc2047 and bnlg1671 as well as bnlg1444 and umc1313, respectively, by linkage analysis. Subsequently, RpiX178-1 was finely mapped between markers SSRZ8 and IDP2347, with genetic distances of 0.6 and 1.1 cM, respectively, and the physical distance of the target region was about 700 kb. RpiX178-2 was also further located between markers bnlg1444 and umc2041, with a genetic distance of 2.4 cM. Moreover, we confirmed that the two genes RpiX178-1 and RpiX178-2 were newly identified and different from those genes known on chrs. 1 and 4 according to results of allelism testing. Herein, we newly identified two genes resistant to P. inflatum, which provided important genetic information for resistance to Pythium stalk rot in maize.

Characteristics of Carbapenem-Resistant Enterobacteriaceae in Ready-to-Eat Vegetables in China.

Vegetables harboring bacteria resistant to antibiotics are a growing food safety issue. However, data concerning carbapenem-resistant Enterobacteriaceae (CRE) in ready-to-eat fresh vegetables is still rare. In this study, 411 vegetable samples from 36 supermarkets or farmer's markets in 18 cities in China, were analyzed for CRE. Carbapenemase-encoding genes and other resistance genes were analyzed among the CRE isolates. Plasmids carrying carbapenemase genes were studied by conjugation, replicon typing, S1-PFGE southern blot, restriction fragment length polymorphism (RFLP), and sequencing. CRE isolates were also analyzed by pulsed-field gel electrophoresis (PFGE). Ten vegetable samples yielded one or more CRE isolates. The highest detection rate of CRE (14.3%, 4/28) was found in curly endive. Twelve CRE isolates were obtained and all showed multidrug resistance: Escherichia coli, 5; Citrobacter freundii, 5; and Klebsiella pneumoniae, 2. All E. coli and C. freundii carried blaNDM, while K. pneumoniae harbored blaKPC-2. Notably, E. coli with blaNDM and ST23 hypervirulent Klebsiella pneumoniae (hvKP) carrying blaKPC-2 were found in the same cucumber sample and clonal spread of E. coli, C. freundii, and K. pneumoniae isolates were all observed between vegetable types and/or cities. IncX3 plasmids carrying blaNDM from E. coli and C. freundii showed identical or highly similar RFLP patterns, and the sequenced IncX3 plasmid from cucumber was also identical or highly similar (99%) to the IncX3 plasmids from clinical patients reported in other countries, while blaKPC-2 in K. pneumoniae was mediated by similar F35:A-:B1 plasmids. Our results suggest that both clonal expansion and horizontal transmission of IncX3- or F35:A-:B1-type plasmids may mediate the spread of CRE in ready-to-eat vegetables in China. The presence of CRE in ready-to-eat vegetables is alarming and constitutes a food safety issue. To our knowledge, this is the first report of either the C. freundii carrying blaNDM, or K. pneumoniae harboring blaKPC-2 in vegetables. This is also the first report of ST23 carbapenem-resistant hvKP strain in vegetables.

High Incidence of Escherichia coli Strains Coharboring mcr-1 and blaNDM from Chickens.

This study investigated the characteristics of Escherichia coli isolates carrying mcr-1-blaNDM from a chicken farm in China. Of the 78 E. coli isolates, 21 clonally unrelated isolates carried mcr-1-blaNDM Diverse IncI2 plasmids disseminated mcr-1, while the dissemination of blaNDM was mediated by diverse IncB/O plasmids. More striking was the colocalization of resistance genes mcr-1 and blaNDM-4 in an IncHI2/ST3 plasmid, which might pose a great challenge for public health.

Emergence of Colistin Resistance Gene mcr-1 in Cronobacter sakazakii Producing NDM-9 and in Escherichia coli from the Same Animal.

We report the presence of mcr-1 in Escherichia coli and carbapenem-resistant Cronobacter sakazakii from the same diseased chicken. The mcr-1 gene linked with ISApl1 was located on two different IncI2 plasmids, including one multidrug plasmid in E. coli, whereas fosA3-blaNDM-9 was on an IncB/O plasmid in C. sakazakii The development of the fosA3-blaNDM-9 resistance region was mediated by IS26 The colocation of mcr-1 or blaNDM-9 with other resistance genes will accelerate the dissemination of the two genes.

Two genes conferring resistance to Pythium stalk rot in maize inbred line Qi319.

Stalk rots are destructive diseases in maize around the world, and are most often caused by the pathogen Pythium, Fusarium and other fungi. The most efficient management for controlling stalk rots is to breed resistant cultivars. Pythium stalk rot can cause serious yield loss on maize, and to find the resistance genes from the existing germplasm is the basis to develop Pythium-resistance hybrid lines. In this study, we investigated the genetic resistance to Pythium stalk rot in inbred line Qi319 using F2 and F2:3 population, and found that the resistance to Pythium inflatum in Qi319 was conferred by two independently inherited dominant genes, RpiQI319-1 and RpiQI319-2. Linkage analysis uncovered that the RpiQI319-1 co-segregated with markers bnlg1203, and bnlg2057 on chromosome 1, and that the RpiQI319-2 locus co-segregated with markers umc2069 and bnlg1716 on chromosome 10. The RpiQI319-1 locus was further mapped into a ~500-kb interval flanked by markers SSRZ33 and SSRZ47. These results will facilitate marker-assisted selection of Pythium stalk rot-resistant cultivars in maize breeding. To our knowledge, this is the first report on the resistance to P. inflatum in the inbred line Qi319, and is also the first description of two independently inherited dominant genes conferring the resistance of Pythium stalk rot in maize.

Identification of the gene Pm47 on chromosome 7BS conferring resistance to powdery mildew in the Chinese wheat landrace Hongyanglazi.

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is an important disease that causes substantial yield losses in wheat (Triticum aestivum) in China and other parts of the world. This foliar disease can be effectively managed by host resistance. The Chinese landrace Hongyanglazi from Shaanxi province is highly resistant to many Bgt isolates at the seedling stage. Genetic analysis using an F2:3 population derived from a cross between Hongyanglazi and susceptible cultivar Zhongzuo 9504 indicated that Hongyanglazi carried a single recessive gene (tentatively designated PmHYLZ) conferring its resistance to Bgt isolate E09. PmHYLZ was flanked by EST marker BE606897 and microsatellite marker Xgwm46 on chromosome 7BS at genetic distances of 1.7 and 3.6 cM, respectively. This gene differed from Pm40, also located on 7BS, by origin, linked markers, and reactions to 13 Bgt isolates. Based on these findings, PmHYLZ was permanently designated as Pm47.