We identified a novel plasmid-mediated colistin resistance gene named mcr-7.1 in K. pneumoniae in China. The prevalence of mcr-7.1 in various species of human and animal origin needs to be investigated immediately.

The mcr-1 gene was detected in 5.11% (58/1136) of Escherichia coli isolates of chicken origin from 13 provinces in China. A novel mcr-1 variant, named mcr-1.3, encoding an Ile-to-Val functional variant of MCR-1 was identified in a sequence type 155 (ST155) strain. An mcr-1.3-containing IncI2 plasmid, pHeN867 (60,757 bp), was identified. The transfer of pHeN867 led to a 32-fold increase in the MIC of colistin in the recipient, exhibiting an effect on colistin resistance that was similar to that of mcr-1. KEYWORDS E. coli, colistin resistance, mcr-1.3, plasmid P olymyxins (polymyxin B and colistin) are a last-resort treatment for infections caused by multidrug-resistant (MDR) Gram-negative bacteria (1). In veterinary use, colistin is administered with food in pig and poultry farming to prevent infections caused by pathogens (2). The mcr-1 gene, which confers plasmid-mediated colistin resistance to Enterobacteriaceae, was identified in an IncI2 plasmid from Escherichia coli and Klebsiella pneumoniae in China in 2016 (3). The mcr-1 gene found in E. coli (4), K. pneumoniae (5), and Salmonella spp. (6) has been proven to disseminate ubiquitously. The transmission of mcr-1-mediated colistin resistance between animals and human has been a threat to human health. It has also been demonstrated that the mcr-1 gene can coexist with bla CTX-M (5), bla NDM (7), and other resistance genes (4), which threatens a return of untreatable infections worldwide. Previous reports described the unique mcr-1 gene sequence compared with that of the originally published sequence (3), which indicates that mcr-1 is relatively conserved. Recently, a point mutation of A¡T at position 8 in mcr-1 was identified in K. pneumoniae (8). To investigate the epidemiology of mcr-1 and its variant, E. coli isolates collected from chickens nationwide in China were assessed.In total, 1,136 nonduplicate E. coli isolates were collected between 2010 and 2015 from sick chickens in 20 provinces and municipalities in China. All of these isolates were preliminarily screened on Mueller-Hinton agar medium with 2 g/ml colistin. Because the cooccurrence of mcr-1 with bla CTX-M may accelerate the transmission of resistance to colistin and cephalosporins, the mcr-1 (3) and bla CTX-M (9) genes were detected by PCR amplification of the isolates with resistance to colistin. The corresponding primers used to amplify the whole mcr-1 gene and parts of the ISApl1 element are listed in Table S1 in the supplemental material. For all of the positive PCR products of mcr-1, Sanger sequencing was performed (Tsingke Biological Technology, Chengdu, China) by using a DNA analyzer (Applied Biosystems, Life Technologies, Carlsbad, CA). We found a total of 58 (5.11%) mcr-1-positive isolates, including one isolate harboring the mcr-1 gene with mutations not found in the originally published gene sequence (3). MICs of colistin

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SXT/R391 integrative and conjugative elements (ICEs) were detected in 8 out of 125 Proteus mirabilis isolates from food-producing animals in China. Whole-genome sequencing revealed that seven ICEs were identical to ICEPmiJpn1, carrying the cephalosporinase gene bla CMY-2 . Another one, designated ICEPmiChn1, carried five resistance genes. All eight ICEs could be transferred to Escherichia coli via conjugation. The results highlight the idea that animal farms are important reservoir of the SXT/ R391 ICE-containing P. mirabilis.

Tibetan Chickens should have unique gastrointestinal microbiota because of their particular habitats. Thus, the aim of this study was to investigate the cecal microbiota of Tibetan Chickens from five typical high‐altitude regions of China. Lohmann egg‐laying hens (LMs) and Daheng broiler chickens (DHs) were chosen as controls. The cecal bacterial populations of Tibetan Chickens were surveyed by high‐throughput sequencing (HTS) of the bacterial 16S rRNA hypervariable region V3‐V4 (16S rRNAV3‐V4) combined with community‐fingerprinting analysis of the 16S rRNA gene based on polymerase chain reaction‐denaturing gradient gel electrophoresis (PCR‐DGGE). The results revealed that the majority of cecal microbiota differed between the Tibetan Chicken and LM/DH. The microbial communities in the cecum were composed of 16 phyla, 28 classes, 36 orders, 57 families, 101 genera, and 189 species. Represented phyla were Bacteroidetes (>47%), Firmicutes (>18.8%), Spirochaetae (>0.3%), and Proteobacteria (>0.4%). Bacteroides and the RC9 gut group were the two most abundant genera. There were relatively more Christensenellaceae, Subdoligranulum, Spirochaeta, and Treponema in Tibetan Chickens, whereas there were more Phascolarctobacterium, Faecalibacterium, Megamonas, and Desulfovibrio in LMs and DHs. The cecal microbiota of Tibetan Chicken have slightly diverged due to exposure to different geographic environments. Differences in the intestinal bacterial communities of Tibetan Chicken and LM/DH were noted.

This study aims at investigating the distribution, antimicrobial resistance, and genetic relationship of Salmonella isolated from 18 farms, their downstream abattoirs, and markets of chickens and pigs in Sichuan province, China. A total of 193 Salmonella isolates were identified from 693 samples with an isolation rate of 26.27% (88/335) in chickens and 29.33% (105/358) in pigs. Salmonella was isolated more frequently in abattoirs and markets than from farms. Serotypes were determined according to the White-Kauffmann-Le Minor scheme and 16 different serotypes were identified, with Derby being the most common, followed by Typhimurium and Meleagridis. Antimicrobial resistance phenotypes and genotypes were studied by using the disk diffusion method and polymerase chain reaction (PCR) amplification, respectively. Overall, 44.04% (n = 85) of all isolates were multidrug resistant (MDR) and resistance to nalidixic acid (51.30%) was the most frequently observed. bla was the most prevalent extended-spectrum β-lactamases gene, and polymyxin resistance gene mcr-1 was present in strains with various serotypes. Multilocus sequence typing indicated that sequence type (ST) had a close relationship with serotype, and 34.20% of all strains were ST40, which was the most prevalent. The unweighted pair group method with arithmetic means (UPGMA) dendrogram of pulsed-field gel electrophoresis showed that Salmonella isolates belonging to the same serovar from different parts of the production chain were highly genetic related, indicating that Salmonella as well as resistance genes could potentially be transmitted from farms to markets. Our study highlights the fact that Salmonella isolates from chicken and pig production chain were frequently exhibiting MDR profiles, and the dissemination of MDR Salmonella from farm to market could pose significant threats to food safety and public health.

Sixteen different sequence types (STs) of Escherichia coli isolates from a commercial swine farm in China were confirmed to coharbor the carbapenem resistance gene bla and the colistin resistance gene mcr-1. Whole-genome sequencing revealed that bla NDM-5 and mcr-1 were located on a 46-kb IncX3 plasmid and a 32-kb IncX4 plasmid, respectively. The two plasmids can transfer together with a low fitness cost, which might explain the presence of various STs of E. coli coharboring bla NDM-5 and mcr-1.KEYWORDS Escherichia coli, carbapenems, bla NDM-5 , colistin, mcr-1, fitness cost C arbapenemase-producing Enterobacteriaceae has become a major public health threat around the world (1). The recently identified carbapenemase New Delhi metallo-␤-lactamase confers resistance to all ␤-lactam antimicrobials except monobactam (2). The NDM-5-encoding gene bla NDM-5 was first identified in an Escherichia coli strain recovered from a patient in the United Kingdom in 2011 (3). Since then, bla was identified in many countries, such as Algeria (4-6), the United States (7), Australia (8), China (9-12), Denmark (13), Japan (14), India (15), and the United Kingdom (3). The widespread occurrence of NDM-5 in recent years should arouse our attention. Colistin is a critically important medication for humans in the treatment of carbapenemaseproducing Enterobacteriaceae, and it has been widely used in veterinary medicine in China (16,17). The first plasmid-mediated colistin resistance gene, mcr-1, was reported in E. coli in 2015 (18). In a short period, colistin-resistant E. coli carrying the mcr-1 gene were reported worldwide (19,20). Recently, mcr-1 was reported to coexist with bla NDM (21-23) and bla CTX-M (24), which brought great challenges for the treatment of bacterial infection. In the present study, we are the first to report the presence of isolates of various sequence types of E. coli coharboring bla NDM-5 and mcr-1 genes from a commercial pig farm in China.A total of 105 anal swabs samples from swine were collected from a commercial pig farm on 1 October 2015 in Sichuan province. E. coli strains were selected by eosinmethylene blue agar, and only 1 isolate was picked up from each sample. All 105 isolates were identified by BD Phoenix 100 diagnostic systems (Sparks, MD). Sixty-four strains were nonsusceptible to imipenem and polymyxin B, identified by the agar dilution method according to Clinical and Laboratory Standards Institute guidelines (25). Isolates were divided into 16 different clones by pulsed-field gel electrophoresis after XbaI digestion according to the standard PulseNet conditions (26) (Fig. 1). Phy-

Anthropogenic activities near urban rivers may have significantly increased the acquisition and dissemination of antibiotic resistance. In this study, we investigated the prevalence of colistin resistant strains in the Funan River in Chengdu, China. A total of 18 mcr-1-positive isolates (17 Escherichia coli and 1 Enterobacter cloacae) and 6 mcr-3-positive isolates (2 Aeromonas veronii and 4 Aeromonas hydrophila) were detected, while mcr-2, mcr-4 and mcr-5 genes were not detected in any isolates. To further explore the overall antibiotic resistance in the Funan River, water samples were assayed for the presence of 15 antibiotic resistance genes (ARGs) and class 1 integrons gene (intI1). Nine genes, sul1, sul2, intI1, aac(6′)-Ib-cr, blaCTX-M, tetM, ermB, qnrS, and aph(3′)-IIIa were found at high frequencies (70–100%) of the water samples. It is worth noting that mcr-1, blaKPC, blaNDM and vanA genes were also found in water samples, the genes that have been rarely reported in natural river systems. The absolute abundance of selected antibiotic resistance genes [sul1, aac(6′)-Ib-cr, ermB, blaCTX-M, mcr-1, and tetM] ranged from 0 to 6.0 (log10 GC/mL) in water samples, as determined by quantitative polymerase chain reaction (qPCR). The sul1, aac(6′)-Ib-cr, and ermB genes exhibited the highest absolute abundances, with 5.8, 5.8, and 6.0 log10 GC/mL, respectively. The absolute abundances of six antibiotic resistance genes were highest near a residential sewage outlet. The findings indicated that the discharge of resident sewage might contribute to the dissemination of antibiotic resistant genes in this urban river. The observed high levels of these genes reflect the serious degree of antibiotic resistant pollution in the Funan River, which might present a threat to public health.