Yeonji Bae

Pathobiological features of a novel, highly pathogenic avian influenza A(H5N8) virus.

The endemicity of highly pathogenic avian influenza (HPAI) A(H5N1) viruses in Asia has led to the generation of reassortant H5 strains with novel gene constellations. A newly emerged HPAI A(H5N8) virus caused poultry outbreaks in the Republic of Korea in 2014. Because newly emerging high-pathogenicity H5 viruses continue to pose public health risks, it is imperative that their pathobiological properties be examined. Here, we characterized A/mallard duck/Korea/W452/2014 (MDk/W452(H5N8)), a representative virus, and evaluated its pathogenic and pandemic potential in various animal models. We found that MDk/W452(H5N8), which originated from the reassortment of wild bird viruses harbored by migratory waterfowl in eastern China, replicated systemically and was lethal in chickens, but appeared to be attenuated, albeit efficiently transmitted, in ducks. Despite predominant attachment to avian-like virus receptors, MDk/W452(H5N8) also exhibited detectable human virus-like receptor binding and replicated in human respiratory tract tissues. In mice, MDk/W452(H5N8) was moderately pathogenic and had limited tissue tropism relative to previous HPAI A(H5N1) viruses. It also induced moderate nasal wash titers in inoculated ferrets; additionally, it was recovered in extrapulmonary tissues and one of three direct-contact ferrets seroconverted without shedding. Moreover, domesticated cats appeared to be more susceptible than dogs to virus infection. With their potential to become established in ducks, continued circulation of A(H5N8) viruses could alter the genetic evolution of pre-existing avian poultry strains. Overall, detailed virological investigation remains a necessity given the capacity of H5 viruses to evolve to cause human illness with few changes in the viral genome.

Pathological Evaluation of Natural Cases of a Highly Pathogenic Avian Influenza Virus, Subtype H5N8, in Broiler Breeders and Commercial Layers in South Korea

SUMMARY Outbreaks of highly pathogenic avian influenza (HPAI) virus, subtype H5N8, were observed in two different flocks of local broiler breeder farms and a commercial layer farm in South Korea. Clinically, the cases were characterized by a gradual increase in mortality, slow transmission, and unrecognizable clinical signs of HPAI. Gross observations in both cases included hemorrhagic or necrotic lesions in internal organs, such as serosal and mucosal membranes, spleen, and pancreas. Both cases exhibited similar histopathologic lesions, including multifocal malacia in the brain and multifocal or diffuse necrosis in the spleen and pancreas. Immunohistochemical results indicated that neurons and glial cells in the brain, myocytes in the heart, acinar cells in the pancreas, and mononuclear phagocytic cells in several visceral organs were immunopositive for avian influenza viral antigen. To experimentally reproduce the low pathogenicity and the mortality observed in these two cases, 18 specific-pathogen-free chickens and 18 commercial layers were divided into an H5N8 virus-inoculated group and a contact-exposed group. The mortality of the chickens in the inoculation group was 50%–100%, whereas the mean time to death was delayed or death did not occur in the contact-exposed group. The distributions of the viral antigens and histopathologic lesions in the experimental study were similar to those observed in the field cases. These findings suggest that the H5N8 virus induces a different pattern of pathobiology, including slow transmission and low mortality, compared with that of other HPAI viruses. This is the first pathologic description of natural cases of H5N8 in South Korea, and it may be helpful in understanding the pathobiology of novel H5N8 HPAI viruses. RESUMEN Reporte de caso- Evaluación patológica de casos clínicos por el virus de la influenza aviar altamente patógeno subtipo H5N8, en reproductores pesados y gallinas de postura comerciales en Corea del Sur. Brotes por el virus de influenza aviar altamente patógena (HPAI), subtipo H5N8, se observaron en dos lotes diferentes de granjas de reproductoras pesadas locales y en una granja de gallinas ponedoras comerciales en Corea del Sur. Clínicamente, los casos se caracterizaron por un aumento gradual de la mortalidad, transmisión lenta y signología no específica de influenza aviar de alta patogenicidad. Las observaciones macroscópicas en ambos casos incluyeron lesiones hemorrágicas o necróticas en los órganos internos, incluyendo membranas serosas y mucosas, el bazo y el páncreas. Ambos casos mostraron lesiones histopatológicas similares, incluyendo malacia multifocal en el cerebro y necrosis multifocal o difusa en el bazo y el páncreas. Los resultados de inmunohistoquímica indicaron reacciones inmunológicas positivas para el antígeno viral de la influenza aviar en las neuronas y células gliales en el cerebro, los miocitos en el corazón, las células acinares del páncreas y en las células mononucleares fagocíticas en varios órganos viscerales. Para reproducir de manera experimental la mortalidad observada en estos dos casos, 18 pollos libres de patógenos específicos y 18 ponedoras comerciales se asignaron en dos grupos, un grupo inoculado con el virus H5N8 y otro grupo de aves expuestas por contacto. La mortalidad de los pollos del grupo inoculado fue del 50% al 100%, mientras que el tiempo promedio de mortalidad se retrasó o no se observó mortalidad en el grupo de aves expuestas por contacto. Las distribuciones de los antígenos virales y las lesiones histopatológicas en el estudio experimental fueron similares a las observadas en los casos de campo. Estos hallazgos sugieren que el virus H5N8 induce un patrón de patobiología diferente, incluyendo una transmisión lenta y baja mortalidad, en comparación con otros virus altamente patógenos de influenza aviar. Esta es la primera descripción patológica de casos clínicos por el subtipo H5N8 en Corea del Sur, y puede ser útil en la comprensión de la biopatología del nuevos virus subtipo de la influenza aviar de alta patogenicidad.

Genetic characterization of three novel chicken parvovirus strains based on analysis of their coding sequences

Chicken parvovirus (ChPV) is one of the causative agents of viral enteritis. Recently, the genome of the ABU-P1 strain of ChPV was fully sequenced and determined to have a distinct genomic composition compared with that of vertebrate parvoviruses. However, no comparative sequence analysis of coding regions of ChPVs was possible because of the lack of other sequence information. In this study, we obtained the nucleotide sequences of all genomic coding regions of three ChPVs by polymerase chain reaction using 13 primer sets, and deduced the amino acid sequences from the nucleotide sequences. The non-structural protein 1 (NS1) gene of the three ChPVs showed 95.0 to 95.5% nucleotide sequence identity and 96.5 to 98.1% amino acid sequence identity to those of NS1 from the ABU-P1 strain, respectively, and even higher nucleotide and amino acid similarities to one another. The viral proteins (VP) gene was more divergent between the three ChPV Korean strains and ABU-P1, with 88.1 to 88.3% nucleotide identity and 93.0% amino acid identity. Analysis of the putative tertiary structure of the ChPV VP2 protein showed that variable regions with less than 80% nucleotide similarity between the three Korean strains and ABU-P1 occurred in large loops of the VP2 protein believed to be involved in antigenicity, pathogenicity, and tissue tropism in other parvoviruses. Based on our analysis of full-length coding sequences, we discovered greater variation in ChPV strains than reported previously, especially in partial regions of the VP2 protein.

Review of Avian Influenza Outbreaks in South Korea from 1996 to 2014

SUMMARY. Since the first outbreak of low pathogenic avian influenza (LPAI) in 1996, outbreaks of LPAI have become more common in Korea, leading to the development of a nationwide mass vaccination program in 2007. In the case of highly pathogenic avian influenza (HPAI), four outbreaks took place in 2003–04, 2006–07, 2008, and 2010–11; a fifth outbreak began in 2014 and was ongoing at the time of this writing. The length of the four previous outbreaks varied, ranging from 42 days (2008) to 139 days (2010–11). The number of cases reported by farmers that were subsequently confirmed as HPAI also varied, from seven cases in 2006–07 to 53 in 2010–11. The number of farms affected by the outbreaks varied, from a low of 286 (2006–07) with depopulation of 6,473,000 birds, to a high of 1500 farms (2008) with depopulation of 10,200,000 birds. Government compensation for bird depopulation ranged from

Eggshell apex abnormalities associated with Mycoplasma synoviae infection in layers

253 million to

The current epidemiological status of infectious coryza and efficacy of PoulShot Coryza in specific pathogen-free chickens.

683 million in the five outbreaks. Despite the damage caused by the five HPAI outbreaks, efficient control strategies have yet to be established. Meanwhile, the situation in the field worsens. Analysis of the five HPAI outbreaks revealed horizontal farm-to-farm transmission as the main factor effecting major economic losses. However, horizontal transmission could not be efficiently prevented because of insufficient transparency within the poultry industry, especially within the duck industry, which is reluctant to report suspicious cases early. Moreover, the experiences and expertise garnered in previous outbreaks has yet to be effectively applied to the management of new outbreaks. Considering the magnitude of the economic damage caused by avian influenza and the increasing likelihood of its endemicity, careful and quantitative analysis of outbreaks and the establishment of control policies are urgently needed.

Pathology and molecular characterization of recent Leucocytozoon caulleryi cases in layer flocks

Eggs exhibiting eggshell apex abnormalities (EAA) were evaluated for changes in shell characteristics such as strength, thickness, and ultrastructure. Mycoplasma synoviae (MS) infection was confirmed by serological assay along with isolation of MS from the trachea and oviduct. Changes in eggshell quality were shown to be statistically significant (p < 0.01). We also identified ultrastructural changes in the mammillary knob layer by Scanning Electron Microscopy. While eggs may seem to be structurally sound, ultrastructural evaluation showed that affected eggs do not regain their former quality. In our knowledge, this is the first report describing the occurrence of EAA in Korea.

Epidemiological Studies of Avian Paramyxovirus Type 4 and 6 in Commercial Chicken Flocks in Korea

Infectious coryza (IC) is an infectious disease caused by Avibacterium (Av.) paragallinarum. IC is known to cause economic losses in the poultry industry via decreased egg production in layers. Between 2012 and 2013, Av. paragallinarum was isolated from seven chicken farms by Chungbuk National University. We identified Av. paragallinarum, the causative pathogen of IC by polymerase chain reaction (PCR) and serovar serotype A, by multiplex PCR. Antibiotic sensitivity tests indicated that a few field-isolated strains showed susceptibility to erythromycin, gentamicin, lincomycin, neomycin, oxytetracycline, spectinomycin, and tylosin. A serological survey was conducted to evaluate the number of flocks that were positive for Av. paragallinarum by utilizing a HI test to determine the existence of serovar A. Serological surveys revealed high positivity rates of 86.4% in 2009, 78.9% in 2010, 70.0% in 2011, and 69.6% in 2012. We also challenged specific pathogen-free chickens with isolated domestic strains, ADL121286 and ADL121500, according to the measured efficacy of the commercial IC vaccine, PoulShot Coryza. We confirmed the effectiveness of the vaccine based on relief of clinical signs and a decreased re-isolation rate of ADL121500 strain. Our results indicate IC is currently prevalent in Korea, and that the commercial vaccine is effective at protecting against field strains.

Amino acid substitutions in low pathogenic avian influenza virus strains isolated from wild birds in Korea

Abstract Leucocytozoonosis was found in three layer farms in chickens with suspected fatty liver or fatty liver hemorrhagic syndrome in Korea between 2009 and 2011. These layer chicken flocks showed both mortality and decreased egg production for one or two weeks when they were between 59 and 82 weeks old. At the necropsy, the most prominent gross lesions were found in the liver, which was enlarged, had a fragile texture, exhibited yellowish discolorations, and had various hemorrhagic lesions. Tissue reactions associated with megaloschizonts specific for Leucocytozoon caulleryi were prominent upon microscopic examination of the liver without significant lipidosis. In addition, the ovaries and uterus were the most affected organs for Leucocytozoon caulleryi multiplication, which led to decreased egg productions. Molecular studies with formalin-fixed, paraffin-embedded tissues were performed in search of a partial region of the cytochrome b gene for hemosporidian parasites. Based on these results, the causal agent was determined to be closely related to Leucocytozoon caulleryi reported in Japan and Malaysia. In this study, we describe recently re-occurring leucocytozoonosis in layer chickens, which required histopathology for disease diagnosis. To prevent outbreaks and maintain chicken health and egg production, layer chickens need to be monitored for symptoms of leucocytozoonosis.

MOLECULAR EPIDEMIOLOGY OF AVIAN POXVIRUS IN THE ORIENTAL TURTLE DOVE (STREPTOPELIA ORIENTALIS) AND THE BITING MIDGE (CULICOIDES ARAKAWAE) IN THE REPUBLIC OF KOREA

Avian paramyxovirus (APMV) type 4 and 6 were isolated during an avian influenza (AI) surveillance program of wild birds. This study also conducted experimental infection of wild-bird-origin APMV type 4 and 6 in specific pathogen free (SPF) chickens to study pathogenicity and transmission within domestic flocks. In addition, serological prevalence data of APMV type 4 and 6 in domestic fowls was conducted with chicken sera collected from 2007 to 2009 in order to understand infection status. The results of the animal experiment showed that APMV type 4 and 6 had the ability to infect chickens with sero-conversion and to transmit the virus from infected birds to contacted birds, but showed low pathogenicity. Serological tests revealed that APMV type 4 was widespread in the poultry industry, especially in layer flocks, but the positive rate for APMV type 6 was very low. This study concluded that wild bird-origin APMV type 4 and 6 could infect the chickens by inter-species transmission and the seroprevalence of APMV type 4 was quite high in Korean poultry. However, since almost all the chicken flocks had a high level of antibody titer against APMV type 1, there was possibility of cross reaction between APMV type 1 and 4, which made the interpretations more complicated. In order to understand infection status in the natural environment, additional study is necessary regarding the seroprevalence of APMV type 4 and 6 in the wild bird population.