Altered gene expression in human brain microvascular endothelial cells in response to the infection of influenza H1N1 virus

Influenza viruses not only cause respiratory illness,but also have been reported to elicit neurological manifestations following acute viral infection.The central nervous system(CNS)has a specific defense mechanism against pathogens structured by cerebral microvasculature lined with brain endothelial cells to form the blood–brain barrier(BBB).To investigate the response of human brain microvascular endothelial cells(hBMECs)to the Influenza A virus(IAV),we inoculated the cells with the A/WSN/33(H1N1)virus.We then conducted an RNAseq experiment to determine the changes in gene expression levels and the activated disease pathways following infection.The analysis revealed an effective activation of the innate immune defense by inducing the pattern recognition receptors(PRRs).Along with the production of proinflammatory cytokines,we detected an upregulation of interferons and interferon-stimulated genes,such as IFN-β/λ,ISG15,CXCL11,CXCL3 and IL-6,etc.Moreover,infected hBMECs exhibited a disruption in the cytoskeletal structure both on the transcriptomic and cytological levels.The RNAseq analysis showed different pathways and candidate genes associated with the neuroactive ligand-receptor interaction,neuroinflammation,and neurodegenerative diseases,together with a predicted activation of the neuroglia.Likewise,some genes linked with the mitochondrial structure and function displayed a significantly altered expression.En masse,this data supports that hBMECs could be infected by the IAV,which induces the innate and inflammatory immune response.The results suggest that the influenza virus infection could potentially induce a subsequent aggravation of neurological disorders.

Association between the Phenotypes and Genotypes of Antimicrobial Resistance in Haemophilus parasuis Isolates from Swine in Quang Binh and Thua Thien Hue Provinces, Vietnam

Haemophilus parasuis (H. parasuis) is one of the bacterial pathogens of great concern as it causes huge economic losses to the swine industry worldwide. One of the reasons why the control of H. parasuis has failed is the increase in antimicrobial resistance (AMR). The country of Vietnam has the second-largest pig production in Asia. However, there is still a lack of data about the AMR prevalence of H. parasuis in Vietnam.The purpose of this study is to investigate the prevalence of AMR and analyze the association between AMR and AMR genes (ARGs). The H. parasuis strains used in this research were isolated from swine in the Quang Binh and Thua Thien Hue Provinces, Central Vietnam, as reported in our previous study. All of the strains were tested for AMR against 25 antibacterial agents using the broth microdilution method and for the presence of ARGs using the polymerase chain reaction (PCR) method. The tested strains were shown to have a high frequency of resistance to trimethoprim/sulfamethoxazole (94.6%), followed by resistance to colistin, chloramphenicol, gentamicin, penicillin, lincomycin, and amoxicillin. The most prevalent ARGs in these strains were blaTEM-1 (94.6%), int (76.8%), gyrA (58.9%), and rmtD (50.0%). Cefuroxime, chloramphenicol, and tobramycin resistances were strongly correlated with the presence of the ARGs blaROB-1 (odds ratio (OR) = 26.3, 95% confidence interval (CI) 2.7–255.7, p = 0.002), catl (OR = 25.1, 95% CI 2.4–258.9, p = 0.004), and strB (OR = 23.5, 95% CI 2.6–212.6, p = 0.001), respectively.This study reveals for the first time the current situation of H. parasuis AMR in Central Vietnam, which is helpful for the clinical control of this disease, as well as for the development of policies and clinical practice guidelines to reduce AMR in swine production in Central Vietnam.

Pasteurella multocida capsular:lipopolysaccharide types D:L6 and A:L3 remain to be the main epidemic genotypes of pigs in China

Pasteurella multocida is a leading cause of respiratory disorders in pigs.This study was designed to understand the genotypical and antimicrobial resistant characteristics of P.multocida from pigs in China.To achieve this,we briefly investigated 158 P.multocida isolates from pigs with respiratory disorders in China between 2019 and 2020.Genotyping through multiplex PCR assays assigned these 158 isolates into capsular genotypes A(60.13%,95/158),D(35.44%,56/158),F(4.43%,7/158),and/or lipopolysaccharide(LPS)genotypes L3(28.48%,45/158)and L6(66.46%,105/158).In addition,eight isolates(5.06%,8/158)were found to be nontypable using the LPS genotyping method.When combining the capsular genotypes and the LPS genotypes,D:L6(34.81%,55/158)and A:L6(31.65%,50/158)were the predominant genotypes,followed by A:L3(24.05%,38/158).PCR detection of virulence factor-encoding genes showed that over 80%of the isolates were positive for exbB,tonB,exbD,ompH,ptfA,fimA,sodA,sodC,fur,ompA,oma87,plpB,hsf-2,nanH and hgbB,suggesting the presence of these genes were broad characteristics of P.multocida.We also found approximately 63.92%(101/158),51.27%(81/158),8.86%(14/158),7.59%(12/158),3.16%(5/158),0.63%(1/158),and 0.63%(1/158)of the isolates grew well in media with the presence of colistin(4μg/mL),tetracycline(16μg/mL),tigecycline(1μg/mL),ampicillin(32μg/mL),chloramphenicol(32μg/mL),cefepime(16μg/mL),and ciprofloxacin(1μg/mL),respectively.This study contributes to the understanding of genotypes and antimicrobial resistance profile of P.multocida currently circulation in pigs of China.

Characteristics of colistin-resistant Escherichia coli from pig farms in Central China

The emergence and dissemination of colistin resistance in Enterobacterioceae mediated by plasmid-borne mcr genes in recent years now pose a threat to public health.In this study,we isolated and characterized colistin-resistant and for mcr-positive£coli from pig farms in Central China.Between 2018 and 2019,594 samples were collected and recovered 445 E.coli isolates.Among them,33 with colistin resistance phenotypes and 37 that were positive for mcr genes were identified,including 34 positive for mcr-1,one positive for mcr-3,and two positive for both mcr-1 and mcr-3.An insertion of nine bases("CTGGATACG")into mcr-7 in four mcr-positive isolates led to gene dysfunction,and therefore did not confer the colistin resistance phenotype.Antimicrobial susceptibility testing revealed that 37 mcr-positive isolates showed severe drug resistance profiles,as 50% of them were resistant to 20 types of antibiotics.Multilocus sequence typing revealed a heterogeneous group of sequence types in mcr-positive isolates,among which ST10(5/37),ST156(5/37),and 5T617(4/37)were the predominant types.Plasmid conjugation assays showed that mcr-carrying plasmids of 25 mcr-positive isolates were conjugated with£coli recipient,with conjugation frequencies ranging from 1.7 × 10^(-6) to 4.1 × 10^(-3) per recipient.Conjugation of these mcr genes conferred a colistin resistance phenotype upon the recipient bacterium.PCR typing of plasmids harbored in the 25 transconjugants determined six types of plasmid replicons,including lncX4(14/25),FrepB(4/25),Incl2(3/25),lncHI2(2/25),FIB(1/25),and Inch(1/25).This study contributes to the current understanding of antibiotic resistance and molecular characteristics of colistin-resistant£coli in pig farms.