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We identified clade 2.3.4.4 highly pathogenic avian influenza A(H5N6) viruses from whooper swans (Cygnus cygnus) found dead in Mongolia. The identification of these infections in wild birds in this area is of concern because of the potential for virus dissemination during fall migration.

Since 2014, H5Nx clade 2.3.4.4 highly pathogenic avian influenza viruses (HPAIV) have caused outbreaks in wild birds and poultry in multiple continents, including Asia, Europe, Africa, and North America. Wild birds were suspected to be the sources of the local and global spreads of HPAIV. This study evaluated the infectivity, pathogenicity, and transmissibility of clade 2.3.4.4 H5N6 HPAIV in mandarin ducks (Aixgalericulata) and domestic pigeons (Columbia livia domestica). None of the birds used in this study, 20 mandarin ducks or 8 pigeons, showed clinical signs or mortality due to H5N6 HPAI infection. Two genotypes of H5N6 HPAIV showed replication and transmission by direct and indirect contact between mandarin ducks. H5N6 HPAIV replicated and transmitted by direct contact between pigeons, although the viral shedding titer and duration were relatively lower and shorter than those in mandarin ducks. Influenza virus antigen was detected in various internal organs of infected mandarin ducks and pigeons, indicating systemic infection. Therefore, our results indicate mandarin ducks and pigeons can be subclinically infected with clade 2.3.4.4 H5N6 HPAIV and transfer the virus to adjacent birds. The role of mandarin ducks and pigeons in the spread and prevalence of clade 2.3.4.4 H5N6 viruses should be carefully monitored.

Highly pathogenic avian influenza (HPAI) is considered as one of the most devastating poultry diseases. It is imperative to immediately report any known outbreaks to the World Organization for Animal Health. Early detection of infected birds is of paramount importance to control virus spread, thus minimizing the associated economic loss. In this study, thermal imaging camera devices were used to detect change in the maximum surface temperature (MST) of chickens (n = 5) and ducks (n = 2) as an early indicator of experimental HPAI infection. The MST of both chickens and ducks increased at least 24 h before the manifestation of clinical signs of HPAI infection, depending on the severity of the infection. The basal MST was recorded for broiler chickens housed under small pen and normal farm conditions without intentional infection. A threshold cutoff of MST was established based on the circadian rhythm of normal MST. This study suggests that thermal imaging of chickens and ducks is a promising tool to screen any potential HPAI-infected flock in order to expedite HPAI diagnosis.

An avian influenza A(H6N5) virus with all 8 segments of North American origin was isolated from wild bird feces in South Korea. Phylogenetic analysis suggests that this virus may have been introduced into Asia by wild birds, highlighting the role of wild birds in the dispersal of these viruses.

We report the first detection of Y280-lineage H9N2 avian influenza viruses in live bird markets in Korea during July 2020. The viruses were isolated from domestic ducks and chickens traded in three markets in two different provinces, indicating dispersal of the newly introduced viruses. Complete genome sequencing and comparative phylogenetic analyses of all eight gene segments of the viruses showed high nucleotide homology to a Y280-lineage H9N2 avian influenza virus isolated in a chicken farm in China, which belongs to one of the most prevalent H9N2 genotypes in China. Increasing human cases of the same genotype H9N2 infection in China and the mammalian specific markers present in the viruses isolated suggest potential implications for public health.

In October 2020, a highly pathogenic avian influenza (HPAI) subtype H5N8 virus was identified from a fecal sample of a wild mandarin duck (Aix galericulata) in South Korea. We sequenced all eight genome segments of the virus, designated as A/Mandarin duck/Korea/K20-551-4/2020(H5N8), and conducted genetic characterization and comparative phylogenetic analysis to track its origin. Genome sequencing and phylogenetic analysis show that the hemagglutinin gene belongs to H5 clade 2.3.4.4 subgroup B. All genes share high levels of nucleotide identity with H5N8 HPAI viruses identified from Europe during early 2020. Enhanced active surveillance in wild and domestic birds is needed to monitor the introduction and spread of HPAI via wild birds and to inform the design of improved prevention and control strategies.

Highly pathogenic avian influenza viruses (HPAIVs) and low pathogenic avian influenza viruses (LPAIVs) represent important threats to the poultry industry and global human health. Due to the high rates of avian influenza virus (AIV) transmission, controlling AIV outbreaks is challenging. HPAIV is known to be transmitted from wild birds to domestic ducks, from which it can be transmitted to layer and broiler chickens. Therefore, surveillance of AIV in domestic ducks and chickens in advance of outbreaks can prevent its spread and enable timely implementation of disease control measures. Certain molecular diagnostic tools can be applied in the field for faster AIV detection. In this study, we evaluated the AIV-detection ability of two insulated isothermal PCR (iiPCR) devices, POCKIT™Micro DUO Nucleic Acid Analyzer (POCKIT DUO) and POCKIT™ Central Nucleic Acid Analyzer (POCKIT Central). We found that the analytical, in vivo and clinical performances of the two POCKIT devices were comparable to those of real-time reverse transcription PCR. Due to their brief protocols and short detection times, POCKIT DUO and POCKIT Central represent promising molecular diagnostic devices for the reliable detection of AIV.

We isolated new reassortant avian influenza A(H5N6) viruses from feces of wild waterfowl in South Korea during 2017–18. Phylogenetic analysis suggested that reassortment occurred between clade 2.3.4.4b H5N8 and Eurasian low pathogenicity avian influenza viruses circulating in wild birds. Dissemination to South Korea during the 2017 fall migratory season followed.

H5Nx clade 2.3.4.4 highly pathogenic avian influenza viruses (HPAIVs) have been disseminated to wide geographic regions since 2014. In 2016, five distinct genotypes (C-1 to C-5) of clade 2.3.4.4c H5N6 HPAIVs were detected in South Korea. In this study, we evaluated the pathogenicity, susceptibility to infection, and transmissibility of the two strains representing the C-1 and C-4 genotypes of the H5N6 viruses, which have different PA and NS gene, in domestic ducks. Although the susceptibility to infection of domestic ducks to the two strains was similar, the C-4 genotype virus induced higher mortality in ducks than C-1 genotype virus. A higher titer of viral shedding were detected in ducks challenged with the C-4 genotype virus compared with the C-1 genotype virus. These results indicated that the reassortment of HPAIVs with prevailing low pathogenic avian influenza viruses could effect on the pathogenicity in ducks.

In recent years, avian paramyxovirus type 4 (APMV-4) frequently isolated from wild and domestic bird populations particularly waterfowls worldwide. However, molecular characteristics and genetic diversity of APMV-4 are uncertain, owing to the limited availability of sequence information. A total of 11 APMV-4 strains from 9850 fecal, swab, and environmental samples were isolated during the surveillance program in wintering seasons of 2013-2017 in South Korea. We performed genetic characterization and phylogenetic analysis to investigate the genetic diversity and relatedness between isolates from the region. We report high APMV-4 genetic diversity (multiple genotypes and sub-genotypes) among wild bird and poultry populations in Korea and that the potential virus exchange occurs between neighboring countries via wild bird migration. Furthermore, our study results suggest the possibility of transcontinental transmission of APMV-4 between Asia and Europe.

The present study was carried out to investigate the effects of stocking density, fumonisin B1 (FB), and mycotoxin binder (TB) on growth performance, bone quality, physiological stress indicators, and gut health in broiler chickens. Day-old Ross 308 male broiler chicks (n = 1,440/experiment) were randomly allocated to 72 floor pens in a 3 × 2 × 2 factorial arrangement, using 3 stocking densities (12.5 birds/m2 [HSD], 10 birds/m2 [MSD], or 7.5 birds/m2 [LSD]), 2 levels of purified FB (0 or 10 ppm), and 2 levels of TB (0 or 0.2%). Each treatment had 6 replicates (n = 6/treatment) and experiments lasted 34 days. All data were analyzed using 3-way ANOVA with stocking density level, FB, and TB as main factors. Body weight gain and feed intake were lower (P < 0.05) in broilers kept at HSD and MSD compared to LSD-housed counterparts. Birds fed an FB-contaminated diet exhibited a higher feed-to-gain ratio compared with those fed an FB-free diet (P < 0.05). None of the treatments affected intestinal morphology or ileal secretory immunoglobulin A levels. Stocking density affected tibia breaking strength (P < 0.05), which was lower in chickens housed at HSD compared with LSD-housed chickens. The heterophil/lymphocyte ratio (H/L ratio) was elevated (P < 0.05) in HSD and MSD groups compared with the LSD group. Serum nitric oxide (NO) levels were elevated (P < 0.05) in chickens fed the FB-contaminated diet compared with the control diet-fed counterparts. Significant interaction (P < 0.05) between FB and TB on serum NO levels was noted. In summary, increasing stocking density lowered growth performance and bone quality, but increased the H/L ratio. Dietary TB did not affect FB-induced increases in the feed-to-gain ratio. No interaction was observed between stocking density and FB for the measured variables.

This study investigated the effects of onion (Allium cepa, L.) extract on the antioxidant activity of lipids in low-and high-fat-fed mouse brain lipids and its structural change during in vitro human digestion. The onion extracts were passed through an in vitro human digestion model that simulated the composition of the mouth, stomach, and small intestine juice. The brain lipids were collected from low- and high-fat-fed mouse brain and then incubated with the in vitro-digested onion extracts to determine the lipid oxidation. The results confirmed that the main phenolics of onion extract were kaempferol, myricetin, quercetin, and quercitrin. The quercetin content increased with digestion of the onion extract. Antioxidant activity was strongly influenced by in vitro human digestion of both onion extract and quercetin standard. After digestion by the small intestine, the antioxidant activity values were dramatically increased, whereas the antioxidant activity was less influenced by digestion in the stomach for both onion extract and quercetin standard. The inhibitory effect of lipid oxidation of onion extract in mouse brain lipids increased after digestion in the stomach. The inhibitory effect of lipid oxidation of onion extract was higher in the high-fat-fed mouse brain lipids than that in the low-fat-fed mouse brain lipids. The major study finding is that the antioxidative effect of onion extract may be higher in high-fat-fed mouse brain lipids than that in low-fat-fed mouse brain lipids. Thus, dietary onion may have important applications as a natural antioxidant agent in a high-fat diet.