Avian metapneumovirus (AMPV) is an emerging pathogen causing respiratory and reproductive illness in poultry worldwide. To demonstrate the presence of AMPV in domestic chickens in Korea, we attempted to isolate AMPV from affected chickens. A cytopathic agent was isolated using chicken tracheal ring culture from dead chickens from a broiler breeder farm with reduced egg production in Korea. This agent, termed SC1509 strain, subsequently passed in Vero cells with distinct cytopathic effects. The SC1509 strain was confirmed as avian metapneumovirus (AMPV) using both RT-PCR test and monoclonal antibody-based immunofluorescence assay. Sequence analysis based on the G glycoprotein revealed that the SC1509 strain had 22.5 to 96.0% nucleotide sequence identity and 11.1 to 92.7% predicted amino acid sequence identity with previously published AMPV strains, particularly with the highest sequence homology (95.8 to 96% for nucleotides and 92.2 to 92.7% for amino acids) to European strains belonging to genotype B. The SC1509 strain was phylogenetically clustered with genotype B viruses, confirming that the SC1509 strain belongs to genotype B. This is the first report of genotype B avian metapneumovirus from chickens in Korea.
Amino Acid Sequence ; Base Sequence ; Chickens ; Fluorescent Antibody Technique ; Genotype ; Glycoproteins ; GTP-Binding Proteins ; Herpesvirus 1, Cercopithecine ; Korea ; Metapneumovirus ; Nucleotides ; Ovum ; Poultry ; Sequence Analysis ; Sequence Homology ; Sprains and Strains ; Vero Cells

BACKGROUND: Three isoforms of the neuronal acetylcholine receptors (AChRs) have been described in muscle epsilonAChRs, gammaAChRs and alpha7AChRs. The adult neuromuscular junctions are composed of epsilonAChRs. During immobilization, new AChRs with subunit compositions of gammaAChRs and alpha7AChRs appear in the perijunctional and extrajunctional area. This study evaluated the contribution of these isoforms to neurotransmission during immobilization, by using waglerin-1 which selectively blocks the epsilonAChRs. METHODS: Male mice (n = 20) were used and each group was divided into sham operated or immobilized. A leg was immobilized in mice for 14 days by pinning, after which nerve-evoked twitch tension was examined under anesthesia in tibialis muscle of both legs, with the contralateral leg serving as control. Neuromuscular transmission was monitored by using a peripheral nerve stimulator with the tibialis muscle and sciatic nerve. Electrical stimuli of 0.2 msec duration were applied to sciatic nerve 2 Hz for 2 sec every 30 sec. After administration of waglerin-1, the evoked twitch was recorded. The percent depression of T1 relative to baseline was transformed to probit scale to determine the effective dose of waglerin-1 for 5%, 50%, and 95% twitch depression. RESULTS: The twitch tension in the control group was higher than immobilized group, confirming the efficacy of immobilization. Waglerin-1 produced 99% twitch suppression on the control group. But on the immobilized group, waglerin-1 produced only 70% twitch suppression even at 37 pg. CONCLUSIONS: The functional role of epsilonAChRs in neurotransmission decreases during immobilization while that of gammaAChRs and alpha7AChRs increases.
Adult ; Anesthesia ; Animals ; Depression ; Hindlimb ; Humans ; Immobilization ; Leg ; Male ; Mice ; Muscles ; Neuromuscular Junction ; Neurons ; Peripheral Nerves ; Protein Isoforms ; Receptors, Cholinergic ; Salicylamides ; Sciatic Nerve ; Synaptic Transmission

BACKGROUND: The precise measurement of body temperature during anesthesia is important to prevent hypothermia.The aim of this study was to compare the urinary bladder temperature to the esophageal, nasopharyngeal, rectal and skin temperatures, and to compare three heating methods during spine surgery. METHODS: Forty-two patients with ASA physical status I-II, who were scheduled to undergo spine surgery in the prone position, were included in this study.The patients were randomly divided into 4 groups:Group I was treated without any heating methods; group 2, with fluid-warmers; group 3, with forced air-warmers; and group 4, with a combination of both heating methods.After the induction of anesthesia, the esophageal, nasopharyngeal, rectal, urinary bladder and skin temperature was monitored every 15 minute for 3 hours.The urinary bladder temperature was compared to the esophageal, nasopharyngeal, rectal and skin temperatures. RESULTS: The urinary bladder temperature was found to be higher than the esophageal and the nasopharyngeal temperatures (P < 0.01).The urinary bladder temperature of group 3 was higher than that of group 1 at 180 minutes after induction of anesthesia (P < 0.05).The urinary bladder temperature of group 4 was higher than that of group 1 at 150 minutes (P < 0.05), as well as at 165 and 180 minutes (P < 0.05).The skin temperatures of groups 3 and 4 were higher than group 1 (P < 0.001). CONCLUSIONS: The urinary bladder temperature was higher than the esophageal temperature and correlated with the esophageal, nasopharyngeal and rectal temperatures.During spine surgery in the prone position, a forced air-warmer was found to be the most effective but a combination of all the methods tested was found to be even more effective.
Anesthesia ; Body Temperature ; Heating ; Hot Temperature ; Humans ; Prone Position ; Skin ; Skin Temperature ; Spine ; Urinary Bladder

Newcastle disease (ND) caused by virulent Newcastle disease virus (NDV) is a highly contagious viral disease of poultry. Virulent NDVs characteristically have a multibasic amino acid sequence (virulence motif) such as (112)RRQKRF(117) at the cleavage site of the precusor fusion (F0) protein. The antigenic and immunogenic characteristics of the virulence motif (112)RRQKRF(117) in the F0 protein of virulent NDVs were investigated. Epitope mapping analysis revealed that a RRQKRF-specific monoclonal antibody 4G2 recognized the KRF section of the motif. A synthetic peptide bearing the RRQKRF motif reacted strongly with sera from virulent NDV (with RRQKRF motif)-infected chickens. These sera also showed reactivity to peptides bearing other virulence motifs ((112)KRQKRF(117), (112)RRQRRF(117) and (112)RRRKRF(117)) but not an avirulence motif ((112)GRQGRL(117)) by ELISA. The synthetic bearing RRQKRF motif reacted with 60% to 91% of sera taken from surviving chickens on ND outbreak farms but not with sera from vaccinated birds, even though most of the sera had antibody to NDV due to vaccination. This indicates that the virulence motif has the potential to differentiate virulent NDV infected birds from vaccinated birds.
Amino Acid Motifs/*immunology ; Amino Acid Sequence ; Animals ; Chickens ; Enzyme-Linked Immunosorbent Assay/veterinary ; Epitope Mapping/veterinary ; Newcastle Disease/*immunology ; Newcastle disease virus/*genetics/pathogenicity ; Poultry Diseases/*immunology/*virology ; Serologic Tests/veterinary ; Viral Fusion Proteins/*genetics/immunology ; Virulence/genetics

Several theories emphasize that aging is closely related to oxidative stress and disease. The formation of excess ROS can lead to DNA damage and the acceleration of aging. Vigna angularis is one of the important medicinal plants in Korea. We isolated vitexin from V. angularis and elucidated the lifespan-extending effect of vitexin using the Caenorhabditis elegans model system. Vitexin showed potent lifespan extensive activity and it elevated the survival rates of nematodes against the stressful environments including heat and oxidative conditions. In addition, our results showed that vitexin was able to elevate antioxidant enzyme activities of worms and reduce intracellular ROS accumulation in a dose-dependent manner. These studies demonstrated that the increased stress tolerance of vitexin-mediated nematode could be attributed to increased expressions of stress resistance proteins such as superoxide dismutase (SOD-3) and heat shock protein (HSP-16.2). In this work, we also studied whether vitexin-mediated longevity activity was associated with aging-related factors such as progeny, food intake, growth and movement. The data revealed that these factors were not affected by vitexin treatment except movement. Vitexin treatment improved the body movement of aged nematode, suggesting vitexin affects healthspan as well as lifespan of nematode. These results suggest that vitexin might be a probable candidate which could extend the human lifespan.
Acceleration ; Aging ; Caenorhabditis elegans* ; Caenorhabditis* ; DNA Damage ; Eating ; Heat-Shock Proteins ; Hot Temperature ; Humans ; Korea ; Longevity ; Oxidative Stress ; Plants, Medicinal ; Superoxide Dismutase ; Survival Rate

To expand the epidemiological understanding of Newcastle disease in Jeju Province, Korea, active surveillance was extensively performed through a virological examination for poultry farms and wild birds in Jeju Province during 2007~2008. Samples (swabs or fresh feces) were collected from a total of 6,485 birds including 6,405 domestic birds (chickens, ducks, pheasants, geese, quails, turkeys, and ostriches) and 80 wild birds. A total of 24 hemagglutinating agents were isolated from domestic birds on fourteen farms including five Korean native chicken, one layer chicken, two broiler chicken, four duck and two pheasant farms. The hemagglutinating agents were all identified as lentogenic NDV based on the reverse transcriptase polymerase chain reaction, sequence analysis of amino acids on the F cleavage site and mean death time in chicken embryos. The F gene-based phylogenetic analysis revealed that the NDV isolates were classified into genotypes 1 or 2 of class II. These lentogenic viruses were closely related to NDV vaccine strains used in Jeju Province. Active surveillance conducted for Newcastle disease indicates no scientific evidence of virulent NDV infection in chickens in Jeju Province, Korea since 2005.
Amino Acids ; Animals ; Birds ; Chickens ; Ducks ; Embryonic Structures ; Geese ; Genotype ; Korea ; Newcastle Disease ; Newcastle disease virus ; Poultry ; Quail ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis ; Turkeys

An attenuated vaccine strain AVR1/08 of Korean respiratory type of infectious bronchitis virus (IBV) was developed by 89th passages of IBV D85/06 strain in chicken eggs. The AVR1/08 strain had higher virus titer at least 20 times (10(1.3)) than the parent virus D85/06 by egg inoculation method. The AVR1/08 strain had a single point mutation (S to Y) at position 56 of spike protein of IBV compared to parent virus IBV D85/06 strain. The mutation was observed consistently at viruses after 47th passage in chicken eggs. The AVR1/08 strain showed no virulence even after 6 passages in chickens and all chickens inoculated induced anti-IBV antibody 14 days after vaccination. The AVR1/08 strain had broad protective efficacy against QX type Korean nephropathogenic virus (Q43/06 strain), KM91 type Korean nephropathogenic virus (KM91 strain) and Korean respiratory virus (D85/06 strain). In contrast, Massachusetts (Mass) type attenuated vaccine strain H120 showed protection of 37.5 to 50% against these three viruses. Our results indicate that the AVR1/08 strain has potential as an attenuated vaccine effective in controlling IBVs circulating in Korea.
Chickens ; Eggs ; Humans ; Infectious bronchitis virus ; Korea ; Massachusetts ; Ovum ; Parents ; Point Mutation ; Sprains and Strains ; Vaccination ; Viral Load ; Viruses

Despite the existence of an active vaccination program, recently emerged strains of nephropathogenic infectious bronchitis virus (IBV) in Korea have caused significant economic losses in the poultry industry. In this study, we assessed the pathogenic and antigenic characteristics of a K-IIb type field strain of IBV that emerged in Korea since 2003, such as Kr/Q43/06. Specific pathogen free 1-week-old chickens exhibited severe respiratory symptoms (dyspnea) and nephropathogenic lesions (swollen kidneys with nephritis and urate deposits) following challenge with the recent IBV field strain. The antigenic relatedness (R value), based on a calculated virus neutralization index, of the K-IIb type field strain and K-IIa type strain KM91 (isolated in 1991) was 30%, which indicated that the recent strain, Kr/Q43/06, is a new variant that is antigenically distinct from strain KM91. This report is the first to document the emergence of a new antigenic variant of nephropathogenic IBV in chicken from Korea.
Animals ; Antigens, Viral ; *Chickens ; Coronavirus Infections/epidemiology/*veterinary/virology ; Infectious bronchitis virus/classification/*pathogenicity ; Korea ; Nephritis/*veterinary/virology ; Poultry Diseases/*virology ; Specific Pathogen-Free Organisms ; Virulence

Despite the intensive vaccination policy that has been put in place to control Newcastle disease virus (NDV), the recent emergence of NDV genotype VII strains in Korea has led to significant economic losses in the poultry industry. We ssessed the ability of inactivated, oil-emulsion vaccines derived from La Sota or Ulster 2C NDV strains to protect chickens from challenge with Kr-005/00, which is a recently isolated Korean epizootic genotype VII strain. Six-week-old SPF chickens were vaccinated once and challenged three weeks later via the eye drop/intranasal route. All vaccinated birds were fully protected from disease, regardless of the vaccine strains used. All vaccinated and challenged groups showed significant sero-conversion 14 days after challenge. However, some vaccinated birds, despite being protected from disease, shed the challenge virus from their oro-pharynx and cloaca, albeit at significantly lower titers than the unvaccinated challenged control birds. The virological, serological, and epidemiological significance of our observations with regard to NDV disease eradication is discussed.
Administration, Intranasal ; Animals ; Chickens ; Cloaca/virology ; Disease Outbreaks/prevention & control/*veterinary ; Korea ; Newcastle Disease/*immunology/prevention & control ; Newcastle disease virus/*immunology ; Ophthalmic Solutions ; Poultry Diseases/*immunology/prevention & control ; *Vaccines, Inactivated/administration & dosage ; Viral Vaccines/*administration & dosage ; Virus Shedding/drug effects

Infectious bursal disease virus (IBDV) causes a highly contagious and immunosuppressive disease of chicken. Agar gel immunodiffusion using IBDV antigen extracted from bursa of Fabricius of infected chicken has been used officially for diagnosis of IBDV in Korea. In this study, in order to replace the IBDV whole virus antigen with non-infectious antigen, recombinant VP2 protein (rVP2) of IBDV was produced using recombinant baculovirus expression system. Purified baculovirus-expressed rVP2 was used as an antigen in an agar gel immunodiffusion (AGID). rVP2 antigen precipitated specifically IBDV antibodies. AGID using rVP2 antigen detected anti-IBDV antibodies from 6 dpi to 28 dpi (termination of the experiment) when specific pathogen free chickens were experimentally infected with IBDV 52/70 strain. This was consistent with result by AGID using IBDV antigen, virus neutralization test (VNT) and a commercial ELISA kit (except for one serum). The sensitivity of rVP2 was the same with that of IBDV antigen when field sera (n=324) were tested by AGID. However, AGID using rVP2 antigen detected maternal antibodies from broiler chickens (n=20) on a broiler farm up to 15 days old, although the detection rate of the AGID was relatively low compared to a commercial ELISA kit. Our results indicate that IBDV whole virus antigen from IBDV infected chickens would be replaced with recombinant VP2 protein as an antigen for AGID.
Agar ; Animals ; Antibodies ; Baculoviridae ; Bursa of Fabricius ; Chickens ; Enzyme-Linked Immunosorbent Assay ; Immunodiffusion ; Infectious bursal disease virus ; Korea ; Neutralization Tests ; Specific Pathogen-Free Organisms ; Sprains and Strains ; Staphylococcal Protein A ; Viruses