Avian infectious bronchitis has caused huge economic losses in the poultry industry. Previous studies have reported that infectious bronchitis virus (IBV) infection can produce cytopathic effects (CPE) and apoptosis in some mammalian cells and primary cells. However, there is little research on IBV-induced immune cell apoptosis. In this study, chicken macrophage HD11 cells were established as a cellular model that is permissive to IBV infection. Then, IBV-induced apoptosis was observed through a cell viability assay, morphological changes, and flow cytometry. The activity of caspases, the inhibitory efficacy of caspase-inhibitors and the expression of apoptotic genes further suggested the activation of apoptosis through both intrinsic and extrinsic pathways in IBV-infected HD11 cells. Additionally, ammonium chloride (NH4Cl) pretreated HD11 cells blocked IBV from entering cells and inhibited IBV-induced apoptosis. UV-inactivated IBV also lost the ability of apoptosis induction. IBV replication was increased by blocking caspase activation. This study presents a chicken macrophage cell line that will enable further analysis of IBV infection and offers novel insights into the mechanisms of IBV-induced apoptosis in immune cells.
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-
Porcine epidemic diarrhea virus (PEDV), which re-emerged in China since 2010, has swept across the whole country leading to tremendous economic losses. In this study, a total of 645 diarrhea samples collected from 156 pig farms in Sichuan and Guizhou province during 2014-2018 were tested for PEDV. We found that samples from 47.66% (84/156) of the farms were positive for PEDV with an overall detection rate of 35.81% (231/645). Fifty-two strains were selected for full-length S gene analyses, and these strains were classified into three subgroups, an S-INDEL subgroup (G1c), and two non-S-INDEL subgroups (G2b, AJ1102-like and G2c), accounting for 15.38% (8/52), 23.08% (12/52) and 59.62% (31/52) of the total analysed strains, respectively. We found these three subgroups of PEDV coexisted in Sichuan province, and the S-INDEL strain was detected in Guizhou. Further antigenic variation analysis of the neutralizing epitopes (S10, COE, SS2, SS6 and 2C10) on the spike protein revealed that the S-INDEL and non-S-INDEL strains shared similar variation features in COE and SS6, but exhibited distinct variation patterns in the S10 domain. Unique variation patterns on N-glycosylation sites in the S protein were also observed for the S-INDEL and non-S-INDEL strains. Moreover, nine strains (three from each subgroup) were subjected to full-genome characterization.Complete genome phylogeny showed an inconsistent tree topology for genotyping, with two G2c strains grouped into the GII-b (AH2012-like) genogroup and the remaining seven strains including three S-INDEL strains grouped into the GII-c genogroup. Further recombination analyses indicated that six of the GII-c strains probably originated from intra-genogroup recombinations. Notably, three newly emerged S-INDEL strains with novel recombination patterns were first identified. Together, our data revealed a new status of PEDV in southwest China, which can increase understanding of the prevalence, genetic characteristics and evolutionary profiles of circulating PEDV strains in China.
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