ABSTRACT
Twelve nucleotides located at the 3′ end of viral genomic RNA (vRNA) are conserved among influenza A viruses (IAV) and have a promoter function. Hoffmann's 8-plasmid reverse genetics vector system introduced mutations at position 4, C nucleotide (C4) to U nucleotide (U4), of the 3′ ends of neuraminidase (NA) and matrix (M) vRNAs of wild-type A/PR/8/34 (PR8). This resulted in a constellation of C4 and U4 vRNAs coding for low (polymerases) and relatively high (all others) copy number proteins, respectively. U4 has been reported to increase promoter activity in comparison to C4, but the constellation effect on the replication efficiency and pathogenicity of reverse genetics PR8 (rgPR8) has not been fully elucidated. In the present study, we generated 3 recombinant viruses with C4 in the NA and/or M vRNAs and rgPR8 by using reverse genetics and compared their pathobiological traits. The mutant viruses showed lower replication efficiency than rgPR8 due to the low transcription levels of NA and/or M genes. Furthermore, C4 in the NA and/or M vRNAs induced lower PR8 virus pathogenicity in BALB/c mice. The results suggest that the constellation of C4 and U4 among vRNAs may be one of the multigenic determinants of IAV pathogenicity.
Subject(s)
Animals , Mice , Clinical Coding , Influenza A virus , Influenza, Human , Neuraminidase , Nucleotides , Orthomyxoviridae , Reverse Genetics , RNA , VirulenceABSTRACT
Necrotic enteritis (NE) caused by Clostridium (C.) perfringens commonly occurs in domestic broiler farms since antibiotic supplementation in poultry feed has been banned. We evaluated the antibacterial activities of medicinal plant extracts against C. perfringens isolates to select alternative compounds for preventing NE. We compared antibacterial activities using two methods and evaluated susceptibilities of the isolates based on minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Two (Fraxinus rhynchophylla Hance [FRH] and Geranium koreanum Kom. [GKK]) of the 30 plant extracts had potent antibacterial activities against C. perfringens ATCC 13124 in two assays. The MIC values for FRH and GKK against 20 C. perfringens isolates were 128~256 microg/mL and 32~128 microg/mL, respectively. The geometric MIC mean values for the two extracts were 147.2 microg/mL and 68.8 microg/mL, respectively. The MBCs for the two extracts against the same strains were 1,024~2,048 microg/mL and 256~1,024 microg/mL, respectively. The geometric mean MIC and MBC for GKK were about two-fold lower than those of FRH. The modified spot-on-lawn assay may be useful for measuring primary antibacterial potential. FRH and GKK are expected to be used as feed additives to prevent or treat NE in veterinary practice.
Subject(s)
Chickens , Clostridium , Clostridium perfringens , Enteritis , Fraxinus , Geranium , Microbial Sensitivity Tests , Plant Extracts , Plants, Medicinal , PoultryABSTRACT
We demonstrated that a mineral aqueous solution (MAS) administered to mice functionally and histologically protected against cisplatin-induced acute renal failure (ARF) and CCl4-induced acute liver failure (ALF). In ARF model, 0.4 and 0.2% MAS decreased mortality and the serum concentrations of blood urea nitrogen (BUN) and creatine in mice. Additionally, 0.4 and 0.2% MAS reduced contraction of distal convoluted tubules and suppressed expression of the proinflammatory cytokines interlukein-6 (IL-6) and tumor necrosis factor (TNF-alpha) in the kidney. In ALF model, 0.4 and 0.2% MAS decreased serum concentrations of alanine aminotransferase and aspartate aminotransferase in mice. Additionally, 0.4 and 0.2% MAS reduced necrotic areas and suppressed expression of IL-6 and TNF-alpha in the liver. These results indicate that a MAS might have protective effects against ARF and ALF.
Subject(s)
Animals , Mice , Acute Kidney Injury , Alanine Transaminase , Aspartate Aminotransferases , Blood Urea Nitrogen , Contracts , Creatine , Cytokines , Interleukin-6 , Kidney , Liver Failure, Acute , Liver , Mortality , Silicon , Tumor Necrosis Factor-alphaABSTRACT
Liver fibrosis is a chronic liver disease and lots of people in Korea are suffered. There are many efforts to find candidates to suppress liver fibrogenesis and several chemical-induced model or bile duct ligation model have been used to research and develop hepatic fibrogenic suppressor. From the previous study about functional effects of turnip which cultivated in Kangha Island, we got the feasibility which turnip might be able to inhibit heptatic fibrogenesis. TAA is a representative hepatic fibrosis inducer, repeated 7-weeks i.p. injection of it results in hepatic fibrosis. We compared the level of hepatic fibrosis in TAA-turnip group, TAA group, and vehicle control group. Nodules-formed by TAA were observed; they were rarely shown in vehicle control group, observed in most area in TAA group, but only shown in periportal regions in TAA-turnip group. These results were confirmed through Masson's trichrom stain; fibrous structures increased in TAA group (fibrosis score: 4) but significantly decreased in TAA-turnip group (fibrosis score: 2-3). In conclusion, we got the result that turnip water extract has a potency to protect TAA-induced hepatic fibrogenesis but it is necessary further study to find its mechanism.
Subject(s)
Bile Ducts , Brassica napus , Fibrosis , Korea , Ligation , Liver , Liver Cirrhosis , Liver Diseases , WaterABSTRACT
We examined 216 Escherichia coli (E. coli) isolated from chickens and environmental specimens from hatcheries between 2005 and 2006 in order to evaluate the epidemiological prevalence of avian pathogenic E. coli (APEC) in Korea tentatively by multiplex PCR. The multiplex PCR which was used as tentative criteria of APEC targets 8 virulence-associated genes; enteroaggregative toxin (astA), increased serum survival protein (iss), iron-repressible protein (irp2), P fimbriae (papC), aerobactin (iucD), temperature-sensitive hemagglutinin (tsh), vacuolating autotransporter toxin (vat), and colicin V plasmid operon (cva/cvi) genes. The number of detected genes could be used as a reliable index of their virulence. It was demonstrated that E. coli strains already typed as APEC always harbor 5 to 8 genes, but non-APEC strains harbor less than 4 genes. Assuming the criteria of APEC is a possession of more than 5 virulenceassociated genes, we discriminated 24 APEC strains among the 216 E. coli strains. Contamination rates of APEC in the field were 31.3% in layers, 14.0% in broilers, 2.7% in broiler breeders, and 0.0% in environmental specimens from hatcheries. The combinational tendency of APEC examined is a fundamental possession of astA, iss and iucD genes and addition of cva/cvi, tsh, vat, and irp2 genes which have a critical importance for virulent traits of APEC. Compared with intravenous chicken challenge or embryo lethality assay, multiplex PCR method could be useful to discriminate APEC rapidly for convenient diagnosis.
Subject(s)
Chickens , Colicins , Embryonic Structures , Escherichia , Escherichia coli , Hemagglutinins , Hydroxamic Acids , Korea , Multiplex Polymerase Chain Reaction , Operon , Plasmids , PrevalenceABSTRACT
Studies on Marek's disease virus (MDV)-unique genes are important for understanding the biological nature of the virus. Based on complete DNA sequence analyses of the MDV genomes, the MDV genomes contain presumably at least five MDV-unique genes, which are commonly conserved among the three MDV serotypes. A recombinant baculovirus that contains the MDV serotype 2 (MDV2)-unique gene, ORF873, under the polyhedrin promoter was constructed and designated rAcORF873. Polyclonal and monoclonal antibodies, which recognize the recombinant MDV2 ORF873 protein in Spodoptera frugiperda clone 9 (Sf9) cells infected with rAcORF873, were prepared by immunizing mice with a recombinant fusion protein expressed in Escherichia coli. Immunoblot analyses with the antibodies revealed a major protein band with a molecular mass of 108-kDa in both MDV2-infected chick embryo fibroblasts (CEF) and rAcORF873-infected Sf9 cells. By indirect immunofluorescence analyses using monoclonal antibody, the authentic ORF873 protein was localized in the cytoplasm of MDV2-infected CEF cells. The monoclonal and polyclonal sera, which were generated in the present study and reacted effectively to MDV2 ORF873 protein, are considered to be useful reagents for further studying the role(s) of the ORF873 protein in MDV2 infection.